NEW: Programme and Abstract Book - Euromar 2009

EUR 

EUROMAR 2009 

Programme and 

Abstract Book 

MAR 

5th – 9th July 2009, Göteborg, Sweden 

www.euromar.org 

Magnetic Resonance Conference

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Programme and Abstract Book 


MAGNETIC RESONANCE CONFERENCE 

5-9 July 2009, Göteborg, Sweden 


3


Magnetic Resonance Conference 

5-9 July, 2009, Göteborg, Sweden 

International Programme Committee 

Mikael Akke, Lund University, Lund 

Lucia Banci, University of Florence, Florence 

Martin Billeter, University of Gothenburg, Göteborg 

Sabine van Dorslaer, University of Antwerp, Antwerp 

Philip Grandinetti, Ohio State University, Columbus 

Astrid Gräslund, Stockholm University, Stockholm 

Robert Griffin, Massachusetts Institute of Technology, Cambridge 

Christian Griesinger, Max Planck Institute, Göttingen 

Torleif Härd, Swedish Agricultural University, Uppsala 

Göran Karlsson, University of Gothenburg, Göteborg 

Malcolm Levitt, University of Southampton, Southampton 

Peter Stilbs, Royal Institute of Technology, Stockholm 

Local Organizing Committee 

Martin Billeter, University of Gothenburg, Göteborg 

Göran Karlsson, University of Gothenburg, Göteborg 

Magnus Nydén, Chalmers University of Technology, Göteborg 

and 

with extensive support from the CONGREX Group. 

4 Euromar Magnetic Resonance Conference

Introduction 

EUROMAR is a well established conference in the field of magnetic resonance research. It builds 

on the history of the European Experimental NMR Conference, The Ampere Congress and the 

UK RSC NMR Discussion Group, meetings which merged in 2005, each with a history that dates 

back several decades. 

The programme committee has put together an exciting programme that reflects the vitality 

and diversity of the magnetic resonance field. The interdisciplinarity, which is a hallmark of 

the EUROMAR conferences, is maintained in the dynamic combination of invited and selected 

presentations. 

Several institutional and private sponsors have contributed to keep the registration fees relatively 

low. With their support, it has been possible for young scientists to register at half the regular fee, 

and numerous grants have also been available. We are very grateful for this support. 

We hope that you will enjoy the science but also the setting of this year’s EUROMAR conference. 

Göteborg is a city of science and knowledge, but also a city with a history that extends out and 

invites other cultures to participate in forming the future. As chairman of this conference and 

on behalf of the organizing committee and the EUROMAR board it is my pleasure to invite you 

to make the EUROMAR 2009 conference in Göteborg an exciting and stimulating meeting, both 

from a personal and a scientific point of view. 

Prof. Göran Karlsson 

Chair of EUROMAR 2009 


5

SUNDAY 5 TH OF JULY 

11:00 Registration 

12:00 - 15:45 User meetings 

Varian Room: Ledningsrummet & Valdemar 

Bruker Room: Runan 

Mestrelab Room: Ascom & Catella 

16:00 Opening of the conference Room: Runan 

Presentation and lectures of the Andrew Prize Winner and the Russell Varian Prize Winner 

19:00 Welcome Reception Room: City Hall 

MONDAY 6 TH OF JULY 

08:30 - 10:00 PLENARY SESSION 

Chairperson: Mikael Akke 

08:30 Visualizing RNA in functional motion using NMR Room: Runan 

Hashim M al-Hashimi. Pl01 

09:15 Focussing on spins Pl02 

Jörg Wrachtrup 

10:00 Coffee 

10:40 – 12:20 PARALLEL SESSIONS 

10:40 - 12:20 Solution NMR Room: Runan 

Chairperson: Astrid Gräslund 

10:40 Limits on control of spin dynamics So01 

Navin Khaneja 

11:20 Prochiral and chiral discrimination in NMR spectra: solutes in variably stretched and compressed So02 

anisotropic gels 

Christoph Naumann; Philip W Kuchel. 

11:40 High resolution NMR in inhomogeneous or fluctuating magnetic fields So03 

Philippe Pelupessy; Enrico Rennella; Geoffrey Bodenhausen 

12:00 Towards real-time NMR spectroscopy of high molecular weight proteins So04 

Carlos Amero; Paul Schanda; Pierre Gans; Bernhard Brutscher; Jerome Boisbouvier 


10:40 - 12:20 Enhanced NMR Room: Palmstedt 

Chairperson: Gil Navon 

10:40 DNP-NMR: Prospects and challenges En01 

Mathilde H Lerche 

11:20 Overhauser polarization of nitroxide radicals in aqueous solutions at high magnetic fields En02 

M. J. Prandolini; M. Gafurov; V. P. Denysenkov; Deniz Sezer; B. Endeward; T. F. Prisner 

11:40 Long-lived states to sustain magnetization enhanced by dynamic nuclear polarization En03 

Paul R Vasos; Riddhiman Sarkar, Ahuja Puneet ; Arnaud Comment; Sami Janin; Jean-Philippe Ansermet; 

Jacques van der Klink; James Konter; Patrick Hautle; Ben van den Brandt; Geoffrey Bodenhausen 

12:00 Reversible interactions with para-Hydrogen enhance NMR sensitivity by polarization transfer En04 

Simon Duckett; Gary Green; Michael Cowley; Kevin Atkinson; Adams Ralph; David Williamson 

12:20 Lunch 

14:00 - 15:00 Poster session 

15:00 - 15:40 Frontiers I-II Room: Runan 

Chairperson: Geoffrey Bodenhausen 

15:00 A microfluidic NMR-chip for the on-line monitoring of a microwave-assisted chemical reaction Fr01 

M. Victoria Gomez; Ángel Díaz-Ortíz; Aldrik Velders; Andrés Moreno; Raluca Fratila; Antonio de la Hoz 

15:20 Nearly 10 6 -fold enhancements in intermolecular 1H double-quantum NMR experiments by nuclear Fr02 

hyperpolarization 

Uzi Eliav; Mor Mishkovsky; Lucio Frydman; Gil Navon 

15:40 Coffee 



Chairperson: Gottfried Otting 

16:20 Escape from flatland: phase-cycling in the third dimension Room: Runan 

Michael Tayler; Andrea Sauerwein; Salvatore Mamone; Michael Knight; Sabrina Marie; Pierre Thureau; Pl03 

Giuseppe Pileio; Marina Carravetta; A. Ganesan; Joern Werner; Malcolm Levitt 

17:05 Paramagnetism in biological solid state and solution NMR Pl04 

Claudio Luchinat 

17:50 Groupment Ampere General Assembly Room: Runan 

18:00 Hospitality Suites - Bruker BioSpin 

7

TUESDAY 7 TH OF JULY 


Chairperson: Sabine van Dorslaer 

08:30 Correlation spectroscopy and distance measurements with high field pulse EPR Room: Runan 

Daniella Goldfarb Pl05 

09:15 Lanthanides for structural biology by NMR Pl06 

Gottfried Otting 

10:00 Coffee 


10:40 - 12:20 EPR Room: Runan 

Chairperson: Gunnar Jeschke 

10:40 Distances and orientations on the nanometer scale from pulsed EPR experiments at high fields Ep01 

Marina Bennati 

11:20 The solvation of small disulfonate anions in water/methanol mixtures characterized by high-field Ep02 

pulse ENDOR 

Dariush Hinderberger; Jeannine Heller 

11:40 Site-directed spin labeling pulse EPR reveals the mechanism of transmembrane signal transfer Ep03 

in halobacterial phototaxis 

Heinz-Jürgen Steinhoff; Meike Döbber; Enrica Bordignon; Johann Klare; Julia Holterhues; Swetlana Martell; 

Nadine Mennes; Lin Li; Martin Engelhard 

12:00 Resolving the 14N and 1H frequencies of the two Cu (II) sites in ascorbate oxidase by high field Ep04 

pulse EPR correlation spectroscopy 

Alexey Potapov; Daniella Goldfarb 

10:40 - 12:20 Imaging Room: Palmstedt 

Chairperson: Bernhard Blümich 

10:40 MRI measurements of fluid flows Im01 

Lynn Gladden 

University of Cambridge, Department of Chemical Engineering, Cambridge, United Kingdom 

11:20 Magnetic Resonance imaging in inhomogeneous fields Im02 

Raphael Paquin; Philippe Pelupessy; Geoffrey Bodenhausen 

11:40 Mapping of drug recrystallization in solid dispersions by 1H and 19F NMR and NMR microimaging Im03 

Carina Dahlberg; Sergey V. Dvinskikh; Michael Schuleit ; István Furó 

12:00 Overview of applications for ultra-low field magnetic resonance Im04 

Henrik Sandin; Yonathan Araya; Mark Flynn; John Gomez; Christina Hanson; Robert Kraus; Per Magnelind; 

Karlene Maskaly; Andrei Matlashov; Pulak Nath; Shaun Newman; Tuba Owens; Mark Peters; Igor Savukov; 

Larry Schultz; Al Urbaitis; Petr Volegov; Vadim Zotev; Michelle Espy 

12:20 Lunch 


14:00 - 15:40 Biomolecules Room: Runan 

Chairperson: Torleif Härd 

14:00 NMR studies of Golgi membrane recognition and proteins in solubilizing nanoparticles Bi01 

Marc Lenoir; Timothy J Knowles; Ünal Coskun; Sabine B. Buschhorn; Jonathan James; Rachael Finka; 

Corinne Smith; Yu-Pin Lin; Tim Dafforn; Kai Simons; Michael Overduin 

14:40 Structure of OmpX in membrane bilayer using NMR Bi02 

Donghan Lee; Korvin F.A. Walter; Ann-Kathrin Brückner; Christian Hilty; Stefan Becker; Christian Griesinger 

15:00 Identification of structure conserving motions in the protein ubiquitin Bi03 

Robert Fenwick; Barbara Richter; Nils Lakomek; Christian Griesinger; Michele Vendruscolo; Xavier Salvatella 


15:20 Expression and isotope labeling of G-protein coupled receptors in E. coli Bi04 

Lada Petrovskaya; Olga Bocharova; Katya Lyukmanova; Zakhar Shenkarev; Vladimir Chupin; Marcel Blommers; 

Alexander Arseniev 

14:00 - 15:40 Small molecules Room: Palmstedt 

Chairperson: Mate Erdelyi 

14:00 Conformational changes and biological activities: From small molecules to large biomolecular complexes Sm01 

Horst Kessler 

14:40 Configurational assignment of complex natural products by NMR and distance geometry; Sm02 

application to pyrrole-imidazole alkaloids 

Matthias Köck ; Gesine Schmidt 

15:00 Conformational study on the tripeptide Hyp(OBn)–β-ACC–Pro–OBn in solution by NMR and MD Sm03 

Matthias Fleischmann; Markus Schmid; Valerio D’Elia; Oliver Reiser; Wolfram Gronwald; Ruth Maria Gschwind 

15:20 Determination of structure and dynamics of an organocatalyst using residual dipolar coupling Sm04 

Christina Thiele; Sven Heiles 

15:40 Coffee 


16:20 - 18:00 Relaxation Room: Runan 

Chairperson: Miquel Pons 

16:20 A unified representation of protein structural dynamics in solution Re01 

Martin Blackledge 

17:00 Investigation of correlated protein motions by the measurement of cross-correlated relaxation rates Re02 

Korvin Walter; Donghan Lee; Nils-Alexander Lakomek; Robert B. Fenwick; Xavier Salvatella; Stefan Becker; 

Christian Griesinger 

17:20 Global anisotropic motion in proteins in the solid state Re03 

Józef R. Lewandowski; Julien Sein; Martin Blackledge; Lyndon Emsley 

17:40 Water dynamics in living bacterial cells and dormant endospores Re04 

Erik Persson; Bertil Halle 

16:20 - 18:00 Solid state physics Room: Palmstedt 

Chairperson: Janez Dolinšek 

16:20 The use of quadrupole NMR spectroscopy for the investigation of intermetallic compounds Sp01 

Frank Haarmann 

RWTH Aachen, Institute of Inorganic Chemistry, Aachen, Germany 

17:00 Magnetic resonance study of detonation nanodiamonds with surface modified by transition metal ions Sp02 

Alexander Panich; Alexander Shames; Vladimir Osipov; Alexander Alexenskii; Alexander Vul’ 

17:20 Low-temperature NMR and quantum theory of dihydrogen endofullerenes Sp03 

Salvatore Mamone; Min Ge; D. Huvonen; U. Nagel; Alain Danquigny; Francesco Cuda; Martin Grossel; 

Yasujiro Murata; Koichi Komatsu; Malcolm Levitt; Toomas Rõõm; Marina Carravetta 

17:40 Antiferromagnetic insulator parent state of Cs3C60 superconductor Sp04 

Peter Jeglic; Alexey Y. Ganin; Yasuhiro Takabayashi; Matthew J. Rosseinsky; Kosmas Prassides; Denis Arcon 

18:30 Hospitality Suites - Varian 


9

WEDNESDAY 8 TH OF JULY 


Chairperson: Thomas Prisner 

08:30 Recent advances of MAS solid-state NMR using perdeuterated peptides and proteins Room: Runan 

Veniamin Chevelkov; Vipin Agarwal; Rasmus Linser; Muralidhar Dasari; Y. Xue; Nikolai R. Skrynnikov; Bernd Reif Pl07 

09:15 The quantum chemistry of zero field splitting: from high spin molecules to distance measurements Pl08 

Frank Neese 

10:00 Coffee 


10:40 - 12:20 Solid state NMR Room: Runan 

Chairperson: Robert Tycko 

10:40 Structural insights into Ure2p full-length prion fibrils by solid-state NMR spectroscopy Sn01 

Antoine Loquet; Luc Bousset; Carole Gardiennet; Yannick Sourigues; Christian Wasmer; Birgit Habenstein; 

Anne Schuetz; Beat H. Meier; Ronald Melki; Anja Böckmann 

11:20 Determination of a molecular torsional angle in the bathorhodopsin photointermediate of Sn02 

rhodopsin by double-quantum solid-state NMR 

Maria Concistre; Neville McLean; Ole G. Johannessen; Giuseppe Pileio; Petra H. M. Bovee-Geurts; 

Richard C. D. Brown; Willem J. De Grip; Malcolm H. Levitt 

11:40 NMR with small magnets and well-defined fields Sn03 

Bernhard Blümich; Federico Casanova; Ernesto Danieli; Juan Perlo 

12:00 Optimal control in NMR spectroscopy: Design your own experiment Sn04 

Z. Tosner; C. Kehlet; A.B. Nielsen; M. Bjerring; J.T. Nielsen; L.A. Straasoe; M.S. Vinding; T. Vosegaard; 

N. Khaneja; S.J. Glaser; N.C. Nielsen 

10:40 - 12:20 Paramagnetic Room: Palmstedt 

Chairperson: Örjan Hansson 

10:40 Structures and dynamics of weak protein complexes studied with paramagnetic NMR tools Pa01 

Marcellus Ubbink; Miguel Prudencio; Monica Vlasie; Peter Keizers; Xingfu Xu; Hanna Lindfors; Mathias Hass; 

Alexander Volkov; Jonathan Worrall; Mark Overhand 

11:20 The motions of domains, as seen by RDCs Pa02 

Edward d’Auvergne; Nicolas Coudevylle; Brent Dorr; Fernando Rodriguez-Castaneda; Benedikt Frank; 

Christophe Fares; Christian Griesinger 

11:40 Solid-state NMR of microcrystalline paramagnetic Cu (II) protein: relaxation-optimized sequences, Pa03 

ultra-fast MAS and structural constraints 

Ségolène Laage; Roberta Pierattelli; Isabella Felli; Ivano Bertini; Joseph R. Sachleben; Stefan Steuernagel; 

Anne Lesage; Lyndon Emsley; Guido Pintacuda 

12:00 Paramagnetic NMR crystallography Pa04 

Guido Pintacuda; Gwendal Kervern; Lyndon Emsley; Olivier Maury 

12:20 Lunch 

14:00 - 15:00 Poster session 

15:00 - 15:40 Frontiers III-IV Room: Runan 

Chairperson: Christian Griesinger 

15:00 Measurement of direct dipolar 1H-1H contacts in magic angle spinning solid-state NMR Fr03 

Rene Verel; Paul Schanda; Matthias Huber; Theofanis Manolikas; Beat H. Meier 

15:20 The structure of outer membrane protein A of Mycobacterium Tuberculosis Fr04 

Peter Teriete; Adrian Kolodzik; Michael Niederweis; Francesca M. Marassi 

15:40 Coffee 



16:20 - 18:00 Molecular interactions Room: Runan 

Chairperson: Mike Williamson 

16:20 NMR studies of proteins and RNAs in the regulation of gene expression Mi01 

Michael Sattler 

17:00 Phosphorylation and membrane association of the natively unfolded c-Src unique domain Mi02 

Yolanda Perez; Pau Bernadó; Marga Gairí; Miquel Pons 

17:20 Characterization of an enzyme reaction by STD NMR spectroscopy Mi03 

Thomas Hackl; Iris G. Fransson; Bernd Meyer 

17:40 Studying protein binding with solid-state NMR: the interaction of an antifreeze protein with ice Mi04 

Ansgar Siemer; Ann McDermott 

16:20 - 18:00 Computational Room: Palmstedt 

Chairperson: Martin Billeter 

16:20 NMR spectroscopy beyond Nyquist limitation by random sampling and Fourier transform processing Co01 

Wiktor Kozminski 

17:00 Ensemble calculations for non-structured proteins from PRE and RDC data: a case study of 8 M Co02 

urea unfolded ubiquitin at pH 2.5 

Jie-rong Huang; Stephan Grzesiek 

17:20 A practical toolbox for NMR solution structure determination in structural proteomics Co03 

Aleksandras Gutmanas; Alexander Lemak, ; Christophe Fares; Bin Wu; Adelinda Yee; Maria Sunnerhagen; 

Cheryl Arrowsmith 

17:40 Fully automated solution NMR structure determination and recent advances in protein solid-state Co04 

NMR structure determination 

Torsten Herrmann 

18:00 Hospitality Suites - Jeol 


11

THURSDAY 9 TH OF JULY 

08:30 - 10:00 PLENARY SESSIONS 

Chairperson: Malcolm Levitt 

08:30 Multidimensional pulsed field gradient NMR: structure and dynamics from microns to nanometres Room: Runan 

Petrik Galvosas Pl09 

09:15 Magnetic relaxation dispersion studies of water in biological systems Pl10 

Bertil Halle; Erik Persson Sunde; Johan Qvist 

10:00 Coffee 


10:40 - 12:20 Metabolomics Room: Runan 

Chairperson: Vladislav Orekhov 

10:40 NMR based metabolomics in non genotoxic carcinogenicity: methods for expanding the Me01 

coverage of the metabolome 

Claire Waterman; Denis Rubtsov; Julian Griffin 

11:20 Robust 1H NMR-based metabolomic responses in fish exposed to different sewage effluents in Me02 

two separate studies 

Linda Samuelsson; Berndt Björlenius; Lars Förlin; Joakim Larsson 

11:40 Real time observation of TCA cycle metabolites by DNP Me03 

Ildefonso Marin-Montesinos; Alessia Lodi; John Carrigan; Martin Saunders; Christian Ludwig; Ulrich Gunther 

12:00 Intramolecular isotope distributions reveal metabolic regulation Me04 

Jürgen Schleucher 

10:40 - 12:20 Transport and diffusion Room: Palmstedt 

Chairperson: Bertil Halle 

10:40 Association of charged entities as observed by diffusion and electrokinetic NMR Td01 

István Furó 

11:20 Fast q-space acquisition to monitor red blood cell shape evolution Td02 

Guilhem Pages; Timothy Larkin; David Szekely; Allan Torres; Philip Kuchel 

11:40 Water exchange over the cell membrane Td03 

Ingrid Åslund; Agnieszka Nowacka; Markus Nilsson; Daniel Topgaard 

12:00 Can a self-diffusion spectrum of water, as measured by a novel NMR technique, reveal any wet secrets? Td04 

Janez Stepišnik; Igor Serša; Aleš Mohoriè 

12:20 Lunch 

14:00 - 14:45 PLENARY SESSIONS 

Chairperson: Göran Karlsson 

14:00 Solid state NMR of unfolded, misfolded, and self-assembled proteins Room: Runan 

Robert Tycko Pl11 

14:45 Closing of the conference 

17:00 Excursion (boat) 

19:00 Conference Dinner 


CONFERENCE INFORMATION 

Venue 

Chalmers Conference Centre, Chalmersplatsen 1. 

Language 

The official working language of the conference is English. 

Exhibition 

An exhibition will be arranged in conjunction with the conference. 

Badges 

Each participant will receive a name badge upon registration. For 

security reasons, all participants are requested to wear or bring their 

badge during all the conference activities. 

Meals 

Coffee, lunch and welcome reception is included in the registration fee. 

Conference and Registration Desk 

The Conference and Registration Desk is located in the foyer at 

the main entrance. On-site registration is available throughout the 

conference. 

Office hours: 

Sunday: 11:00-18:30 

Monday-Wednesday: 08:00-08:30, 18:00-18:30 and during 

intermissions 

Thursday: 08:00-08:30, 15:00-16:00 and during intermissions. 

SOCIAL EVENTS 

Welcome Reception 

Sunday 5 July at 19.00 – 20.00 

Welcome reception at City Hall - Börsen, hosted by the City of Göteborg 

and The Region of Västra Götaland. 

Free of charge but must be pre-booked on the registration form to 

obtain a ticket. 


Posters 

Posters abstracts are sorted in the same categories as the scientific 

sessions. There are two formal poster sessions, on Monday and 

Wednesday afternoons, from 14:00-15:00. Authors are asked to 

present odd-numbered posters on Monday and even-numbered posters 

on Wednesday. 

The posters can be mounted from 11.00 on Sunday and should be 

removed before 15:30 on Thursday afternoon. The Congress staff 

will remove posters not taken down on time. However, the Congress 

Organizers cannot ensure any further responsibility for the material. The 

poster boards are numbered in the same way as the poster abstracts, 

i.e. Bi05, and grouped according to session. 

Speakers 

Speakers are asked to bring their presentations to the *Valdemar’ room 

well in advance of each session. Stewards are available to assist in 

transferring presentations or setting-up of personal computers. 

Internet 

WiFi is available during the conference. Due to internet security, 

individual log-in (username & password) has to be used. This 

information is available at the conference desk. 

Conference Dinner 

Thursday July 9 at 17.00 – 24.00 

Conference dinner at Älvsborgs Fortress starting with a 1 hour boat 

tour. The boat will leave from Stenpiren (see map) at 17.00 and take 

you on a boat trip in the archipelago for approx. 1 hour. Älvsborgs 

Fortress has an astonishing history starting in the 17th Century until 

modern time. On three occasions the Danish stood at the gate with no 

good attentions – luckily they had, each occasion, to sail away without 

any success. During the time that the fortress was used as a prison, 

several of the most fearful criminals spent their time on the island. In 

the late 19th Century the fortress became an attraction for the residents 

of Gothenburg where they went to breathe fresh air, smoke cigars and 

drink the traditional Swedish liqueur Punsch 

Price per person: SEK 450 

Must be pre-booked on the registration to obtain a ticket. 

13

GENERAL INFORMATION 

Local Transportation 

Chalmers Conference Centre is located in the city centre with walking 

distance to most hotels and shopping areas. If you would like to 

use the tram take No. 6, 7 or 8 who stops outside the entrance 

(Chalmersplatsen). Please note when travelling by tram you have to 

buy a card valid for trams and buses (except Airport buses) in kiosks 

as Pressbyrån and 7-Eleven. Ask the clerk for a suitable ticket for your 

visit. You can also use the ticket machine inside the trams (2 tickets 

within the city centre) where you pay with Swedish coins or in some 

cases Visa or MasterCard. Please note that you are not able to buy a 

ticket from the chauffeur. 

Taxi 

We recommend Taxi Göteborg, VIP-taxi or Taxi Kurir. Ask for fixed price. 

Tourist Information 

The conference Secretariat will be available to give you more 

information about Göteborg, make restaurant reservations, or assist you 

in any other way during your stay. For additional information please visit 

www.goteborg.com 

Banks 

Banks are open between 10.00 and 15.00 on weekdays. Some banks 

in the city centre are open from 09.00 to 17.00. 

Currency 

The official currency is Swedish Krona (SEK). 

1 EUR = SEK 10,86 (June 2009) 

1 USD = SEK 7,83 (June 2009) 

For update on exchange rates please visit www.oanda.com 

Electricity 

Electrical current in Sweden is 220 V/50 Hz. Round, European-style 

two-pin plugs are used. Appliances designed to operate on 110/120 

Volts need a voltage converter and a plug adapter. 

Time Zone 

The time zone in Stockholm is GMT + 1 hour. Daylight Saving Time is 

used during the summer. 

Smoking Policy 

Sweden has a ban on smoking in bars and restaurants. Designated 

smoking areas are located outside. 

Disclaimer 

The Organising Committee and Congrex Sweden AB accept no liability 

for any injuries/losses incurred by participants and/or accompanying 

persons, nor loss of, or damage to, any luggage and/or personal 

belongings. 

Phone Numbers 

Taxi Göteborg +46 (0)31 65 00 00 

SOS 112 

Contact: 

Congrex Sweden AB 

Ref. EUROMAR 2009 

Box 5078, 402 22 Göteborg, Sweden 

Email: euromar2009@congrex.com 

Phone: +46 31 708 60 00 

Fax: +46 31 708 60 25 


Plenary 

Pl01 

Visualizing RNA in functional motion using NMR 

Al-Hashimi, Hashim M. 

University of Michigan, Department of Chemistry & Biophysics, Ann 

Arbor, United States 

RNA molecules process input cellular signals by undergoing large 

conformational changes through mechanisms that remain poorly 

understood. It remains unclear whether cellular signals, ranging 

from small molecules to proteins, induce formation of new RNA 

conformations via “induced-fit” or instead bind and stabilize preexisting 

conformation from a dynamical ensemble via “conformational 

selection”. We present new developments in NMR methodology that are 

providing new insights into these mechanistic aspects of RNA structural 

transitions by making it possible to characterize unbound RNA structural 

ensembles at atomic resolution. Using these new NMR methods, we 

uncover universal spatially structured RNA flexibility that is encoded 

by steric interactions. We provide strong evidence that conformational 

changes involving changes in the orientation of helical domains occur 

by conformational selection and not induced-fit. The atomic-resolution 

unbound RNA structural ensemble permit accurate virtual screening 

against small molecules using computational docking, thus overcoming 

the problem of being able to predict RNA adaptation upon target 

recognition. 

Pl02 

Focussing on spins 

Wrachtrup, Jörg 

University of Stuttgart, Stuttgart, Germany 

Microscopy has revolutionized our understanding of nature. In particular 

optical microscopy has made tremendous contribution to structure 

analysis on the nanometer scale in life science. Yet, if it comes to 

the fine details of e.g. molecular structures it is mostly spectroscopy 

providing the details. In particular techniques based on spins like 

e.g. nuclear- and electron spin magnetic resonance, have provided 

unprecedented insight into the details of complex structures like 

proteins. The talk will describe the combination of both techniques 

leading to unsurpassed sensitivity in magnetic resonance. In particular 

I shall describe the development of a novel high resolution, spin based 

magnetometer with the potential to imagine single spins with nm 

resolution. 

Pl03 

Escape from flatland: phase-cycling in the third dimension 

Tayler, Michael 1 ; Sauerwein, Andrea 1 ; Mamone, Salvatore 1 ; Knight, 

Michael 2 ; Marie, Sabrina 1 ; Thureau, Pierre 1 ; Pileio, Giuseppe 1 ; 

Carravetta, Marina 1 ; Ganesan, A. 1 ; Werner, Joern 2 ; Levitt, Malcolm 1 

1 University of Southampton, School of Chemistry, Southampton, United 

Kingdom; 2 University of Southampton, School of Biological Sciences, 

Southampton, United Kingdom 

Phase-cycling is one of the most powerful techniques in NMR pulse 

sequence design. In conventional phase cycling, NMR signals are 

filtered according to their history of coherence orders - a classification 

of density operator components according to their rotational symmetry 

about a single fixed axis, namely the main magnetic field. Conventional 

phase cycling therefore operates in “flatland” (the plane perpendicular 

to the magnetic field). 

We have explored new types of phase cycle that “escape from 

flatland” by exploiting the rotational symmetry of the density operator 

components in all three rotational directions, not just in the plane 

perpendicular to the magnetic field. The general method is called 

Spherical Tensor Analysis (STA). We have used STA in two new contexts. 

1. We have designed a two-dimensional correlation experiment 

in solution NMR that generates very similar results to the doublequantum-filtered 

COSY experiment, but with twice the signal strength. In 

some cases, not only is the signal twice as strong, but fewer transients 

are needed to collect the data set. Preliminary results have been 

obtained at the time of abstract submission. There are good prospects 

for extending the method to triple-quantum-filtered COSY and E.COSY, 

also with advantages of improved signal strength. 

2. We have explored the use of STA in solid-state NMR experiments for 

estimating internuclear distances. The main motivation is as follows: 

by extracting several spherical signal components from the same data 

set, it is possible to account for relaxation losses, and obtain accurate 

geometrical information even when the coherence transfer curves 

display no oscillatory structure. Preliminary results look quite promising. 

Prospects and limitations of STA will be discussed. 

I also intend to discuss further developments in the area of longlived 

nuclear singlet states, including extremely low-frequency NMR 

experiments in low magnetic field, and the influence of molecular 

geometry on nuclear singlet lifetimes. 

Programme and Abstract Book 15 

Pl04 

Paramagnetism in biological solid state and solution NMR 

Luchinat, Claudio 

University of Florence, CERM, Florence, Italy 

Exploitation of paramagnetism in metalloproteins is largely based 

on relaxation and/or on the combination of the various pieces of 

information derived from the anisotropic magnetic susceptibility tensor 

(pseudocontact shifts and residual dipolar couplings) to learn about 

the relative degrees of freedom of one protein domain with respect 

to another. Broadly speaking, NMR is in principle able to provide 

information on unstructured or partially structured protein systems, 

thereby complementing other structural techniques. In parallel, there 

is a continuing interest in understanding the dynamics of proteins that 

perform their function by changing their structure. The presence of a 

paramagnetic metal ion helps acquiring information on, e.g., global 

order parameters of one domain with respect to another, or on the 

relative population of different conformers. Paramagnetic effects may 

also provide detailed information on modest structural differences that 

may occur in multidomain proteins between solid state and solution. 

Finally, paramagnetic effects have been recently shown to be a very 

promising tool for the determination of protein structures by solid state 

NMR. Indeed, Curie relaxation, often the major source of paramagnetic 

line broadening, is absent in the solid state. Pseudocontact shifts are 

measured as easily as in solution, while distance restraints of NOE 

type are much less readily obtained. As a consequence, the relative 

importance of pseudocontact shifts as structural restraints is higher in 

the solid state than it is in solution. Furthermore, when the magnetic 

susceptibility tensor is strongly anisotropic, the pseudocontact shifts 

in microcrystalline materials may reach out neighboring molecules in 

the crystal, permitting the obtainment of information on the reciprocal 

disposition of the molecules. 

ABSTRACTS PLENARY

ABSTRACTS PLENARY 

Pl05 

Correlation spectroscopy and distance measurements with high 

field pulse EPR 

Goldfarb, Daniella 

Weizmann Institute of Science, Chemical Physics, Rehovot, Israel 

Pulse EPR techniques are routinely used to derive structural information 

on biomolecules. These are based on the measurements of ligand 

hyperfine interactions in paramagnetic metal ion sites in metalloproteins 

and electron spin-spin dipolar interactions in doubly spin labelled 

biomolecules. We present new developments in these two directions 

afforded by our high power high field (W-band, 95 GHz) pulse EPR 

spectrometer. A popular and effective method for resolving and 

determining ligand hyperfine couplings is the 2D HYSCORE (hyperfine 

sublevel correlation) experiment that correlates nuclear frequencies 

belonging to different electron spin manifolds. It is most efficient when 

the hyperfine interaction and the nuclear Zeeman interactions have 

comparable magnitudes and a large enough microwave B1 field is 

essential as both the allowed and forbidden transitions should be within 

the microwave pulses’ bandwidth. These measurements are commonly 

carried out at X-band frequencies (~ 9 GHz), where for low nuclei, like 

14N and 17O, the accessible hyperfine couplings are rather small (< 7 

MHz). For larger hyperfine couplings (10-25 MHz), higher frequencies 

are required. HYSCORE measurements at high frequencies (high fields), 

however, have been so far held back by power limitations. With our 

new power capabilities such measurements became possible and 

several applications will be presented (nitrosyl heme complexes in nitrite 

reeducates, type I copper in ascorbate oxidase and reduced polyoxometalates). 

Distance measurements are usually carried out between 

nitroxide spin labels by pulse double resonance experiments at X-band 

and the accessible distance range is ~1.5-8.0 nm. To increase the 

sensitivity of distance measurements, in terms of sample concentration 

and amount, we explored the possibility of using Gd 3+ (S=7/2) based 

spin labeling for distance measurements at W-band. Several examples 

will be presented and the advantages/disadvantages will be discussed. 

Pl06 

Lanthanides for structural biology by NMR 

Otting, Gottfried 

The Australian National University, Research School of Chemistry, 

Canberra, Australia 

Paramagnetic lanthanide ions present outstanding opportunities to 

accelerate structural biology studies by nuclear magnetic resonance 

(NMR) spectroscopy.1,2 In particular, pseudocontact shifts (PCS) from 

lanthanide labelled proteins provide a route to rapid 3D structure 

determinations of protein-protein complexes. PCS also provide 

quick access to structures of small molecules as they are bound to 

lanthanide-labelled proteins, which is of great interest in rational drug 

design. Other applications include the NMR resonance assignments of 

proteins of known three-dimensional structure and the determination of 

protein folds from a minimal set of structure restraints. New reagents 

will be presented to achieve site-specific attachment of lanthanide ions 

by covalent or non-covalent bonding. They can also be used as reagents 

for weak alignment of proteins in a magnetic field and measurement of 

residual dipolar couplings. 

References 

1. Pintacuda, G., John, M., Su, X.C., Otting, G. (2007) NMR structure 

determination of protein-ligand complexes by lanthanide labeling. Acc. 

Chem. Res. 40, 206-212. 

2. Otting, G. (2008) Prospects for lanthanides in structural biology by 

NMR. J. Biomol. NMR 42, 1-9. 

16 

Pl07 

Recent advances of MAS solid-state NMR using perdeuterated 

peptides and proteins 

Chevelkov, Veniamin 1 ; Agarwal, Vipin 1 ; Linser, Rasmus 1 ; Dasari, 

Muralidhar 1 ; Xue, Y. 2 ; Skrynnikov, Nikolai R. 2 ; Reif, Bernd 1 

1 Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany; 

2 Purdue University, Department of Chemistry, W. Lafayette, United States 

Successful spectral assignment and determination of structural 

constraints in MAS solid-state NMR in isotopically enriched materials 

(mostly 13 C, 15 N) is limited by resolution and sensitivity. We suggest to 

employ perdeuteration and back-substitution of exchangeable protons 

in peptides and proteins in order to reduce 1 H, 1 H dipolar interactions. 

This way, an increase by a factor of 5-9 in sensitivity can be achieved 

compared to the heteronuclear detected version of the experiment. Use 

of higher levels of deuteration yields a further decrease in the observed 

proton line width, allowing to resolve 1 J NH scalar couplings in the proton 

as well as nitrogen dimension. Addition of complexed paramagnetic 

ions reduces drastically the recycle delay in the experiment, while 

the favorable spectral resolution is retained. Similarly, high sensitivitiy 

and high resolution spectra are obtained for methyl containing side 

chains by either incorporating specifically protonated precursors or by 

making use of the residual protons in otherwise perdeuterated proteins 

(HANAH). In addition, we show that deuteration has a beneficial effect 

on the spectral properties of amyloidogenic peptides and membrane 

proteins. We present relaxation experiments to characterize backbone 

and side chain dynamics in the solid-state. We find that relaxation 

parameters in the solid-state and in solution are highly similar. This 

opens promising perspectives for the characterization of protein 

molecular motion in general, as no upper bounds for correlation times 

are imposed by molecular tumbling in the solid-state. 

Pl08 

The quantum chemistry of zero field splitting: from high spin 

molecules to distance measurements 

Neese, Frank 

Universität Bonn, Wegelerstr. 12, Bonn, Germany 

Systems with more than unpaired electron show the phenomenon of 

zero-field splitting. This term in the phenomenological spin Hamiltonian 

is the most important quantity that characterizes high-spin systems. 

Understanding an predicting the zero-field splitting correctly on the 

basis of quantum chemical methods has been an important objective 

of our research in the past few years.[1-8] A number of wavefunction 

based ab initio as well as density functional theory methods to predict 

the ZFS are implemented in the ORCA quantum chemistry program that 

is freely available (http://www.thch.uni-bonn.de/tc/orca/). Important 

applications concern the calculation of the “magnetic anisotropy” in 

single molecule magnets or distance measurements using pairs of spin 

labels. The lecture will present the fundamental theory of the zero-field 

splitting interaction, will discuss the ZFS in transition metal complexes 

and will finally turn to the reliability of the point dipole model for 

estimating distances from EPR or ELDOR measurements. 

[1] Ganyushin, D.; Neese, F. (2006) J. Chem. Phys., 125, 024103 

[2] Neese, F. (2006) J. Am. Chem. Soc., 128, 10213-10222 

[3] Sinnecker, S.; Neese, F. (2006) J. Phys. Chem. A, 110, 12267- 

12275 

[4] Neese, F. (2007) J. Chem. Phys., 127, 164112 

[5] Sander, W.; Grote, D.; Kossmann, S.; Neese, F. (2008) J. Am. Chem. 

Soc., 130, 4396-4403 

[6] Duboc, C.; Collomb, M.-N.; Pecaut, J.; Deronzier, A.; Neese, F. 

(2008) Chem. Eur. J., 21, 6498-6509 

[7] Cirera, J.; Ruiz, E.; Alvarez, S.; Neese, F.; Kortus, J. (2009) Chem. 

Euromar Magnetic Resonance Conference

Eur. J., in press 

[8] Riplinger, C.; Kao, J.P.Y.; Rosen, G.M.; Kathirvelu, V.; Eaton, G.R.; 

Eaton, S.S.; Kutateladze, A.; Neese F. (2009) J. Am. Chem. Soc., in 

press 

Pl09 

Multidimensional pulsed field gradient NMR: structure and 

dynamics from microns to nanometres 

Galvosas, Petrik 

MacDiarmid Institute for Advanced Materials and Nanotechnology, 

Victoria University of Wellington, Wellington, New Zealand 

Pulsed magnetic field gradient NMR, as introduced by Stejskal and 

Tanner [1], has shaped new NMR methodologies for over four decades. 

Quite apart from the vast, and rapidly developing field of MR imaging, 

we are nowadays presented with a whole toolbox of NMR experiments 

that investigate molecular diffusion and flow. 

Advances in pulsed field gradient NMR have been driven by pulse 

sequence sophistication, new data analysis methods and hardware 

development. An example of the latter advance is the use of intense 

magnetic field gradients to observe small molecular displacements, 

thus probing space on a length scale down to some ten nanometres. 

Methods based on pulse sequence sophistication and novel data 

analysis reveal molecular exchange or local anisotropy by means of 

a multidimensional NMR data set and its subsequent inverse Laplace 

transformation. 

Such an array of experiments allows one to extract the dynamic 

properties of complex fluids and their interactions with the environment. 

Furthermore, in the case of porous materials, properties of the pore 

space can be derived from the diffusional behaviour of guest molecules. 

This lecture will present an overview of selected methods along with 

recent examples illustrating their use. These include the molecular 

exchange of polymers in dispersions of hollow polyelectrolyte multilayer 

capsules [2,3], the diffusional mechanisms and self-assembly of 

triblock copolymers in aqueous solution [4], the diffusional behaviour of 

hydrocarbons in zeolite NaX [5] or the anisotropy of diffusion as found in 

plant tissue [6]. 

[1] E. O. Stejskal and J. E. Tanner, J. Chem. Phys. 42, 288 (1965). 

[2] Y. Qiao, P. Galvosas, T. Adalsteinsson, M. Schönhoff and P. T. 

Callaghan, J. Chem. Phys. 122, 214912 (2005). 

[3] R. P. Choudhury, P. Galvosas and M. Schönhoff, J. Phys. Chem. B 

112, 13245-13251 (2008). 

[4] K. Ulrich, P. Galvosas, J. Kärger and F. Grinberg, Phys. Rev. Lett. 102, 

037801 (2009). 

[5] M. Gratz, M. Wehring, P. Galvosas and F. Stallmach, Microporous 

Mesoporous Mat. in Press (2009). 

[6] Y. Qiao, P. Galvosas and P. T. Callaghan, Biophys J. 89, 2899-2905 

(2005). 

Pl10 

Magnetic relaxation dispersion studies of water in biological 

systems 

Halle, Bertil; Persson Sunde, Erik; Qvist, Johan 

Lund University, Lund, Sweden 

Proteins and other biological macromolecules are tightly coupled 

to the surrounding aqueous hydrogen-bond network. This coupling 

drives protein folding and modulates biomolecular recognition and 

conformational dynamics. In addition, internal water molecules are an 

integral part of most proteins, playing key structural, and sometimes 

functional, roles. 

Magnetic relaxation dispersion (MRD) of the water isotopes 1H, 2H 

and 17O is a versatile method for probing the dynamics of water 

molecules interacting with biomolecules in solution as well as in solids. 

MRD studies of water in vivo are also feasible and can shed light on 

fundamental issues in cell biology and elucidate new modes of image 

contrast in tissue MRI. 

In this lecture, I will briefly review the MRD method and illustrate its 

power by some recent applications to proteins. 


Pl11 

Solid state NMR of unfolded, misfolded, and self-assembled 

proteins 

Tycko, Robert 

National Institutes of Health, Bethesda, United States 

Solid state NMR methods are particularly valuable when applied to 

systems and questions that can not be addressed adequately by other 

experimental techniques. This obvious statement will be illustrated by 

several examples from our recent work: (i) Relatively little is known 

about the molecular structural details of proteins in unfolded and 

partially folded states. We have used solid state NMR methods to 

determine backbone conformational distributions at specific sites 

in frozen solutions of the model protein HP35, unfolded by addition 

of GdnHCl. We have also developed a method for rapid freezing of 

protein solutions, on the 5-10 µs time scale, that allows us to trap and 

characterize intermediate states in the folding of HP35 from an initial 

thermally unfolded state; (ii) Our studies of amyloid fibrils, especially 

fibrils formed by the β-amyloid peptide (Aβ) associated with Alzheimer’s 

disease, reveal a surprisingly high degree of structural polymorphism, 

raising the question of which Aβ fibril structures are most relevant 

to disease. Recent experiments on a disease-associated mutant of 

Aβ (D23N, or Iowa mutant) demonstrate the existence of a surprising 

antiparallel β-sheet structure that has not been observed in wild-type 

Aβ fibrils. Solid state NMR measurements on Aβ fibrils derived from 

human brain tissue suggest that a specific polymorph develops in 

Alzheimer’s disease; (iii) Initial studies of tubular structures formed 

by spontaneous self-assembly of the HIV-1 capsid protein suggest 

that solid state NMR will provide new information about the structures 

of viral capsids in asymmetric viruses. New methods (i.e., pulse 

sequences) and technologies (e.g., ultra-low-temperature magic-angle 

spinning) that contribute to these projects will also be described. 

ABSTRACTS PLENARY

ABSTRACTS ORAL 

Fr01 

18 

Frontiers 

A microfluidic NMR-chip for the on-line monitoring of a 

microwave-assisted chemical reaction 

Gomez, M. Victoria 1 ; Díaz-Ortíz, Ángel 1 ; Velders, Aldrik 2 ; Moreno, 

Andrés 1 ; Fratila, Raluca 2 ; de la Hoz, Antonio 1 

1 University of Castilla-La Mancha, Facultad de Quimicas, Ciudad Real, 

Spain; 2 University of Twente, Supramolecular Chemistry and Technology, 

Enschede, Netherlands 

The miniaturization of the detection radiofrequency coil is an interesting 

approach in NMR to overcome its intrinsically low sensitivity, since the 

signal to noise ratio is inversely proportional to the coil diameter [1]. 

Different microcoil geometries, planar, solenoidal and stripline, can be 

used, allowing the analysis of mass-limited and volume-limited samples 

with an improvement of sensitivity compared to the conventional NMR 

probe. 

Planar microcoils [2] are easier to integrate in microfabrication 

processes using photolithography techniques allowing a precisely 

controlled geometry and an accurate coil sample positioning. These 

microcoils can be implemented in a microfluidic system, resulting a 

flow-NMR (micro)probe which can be hyphenated to other techniques. 

Microwave-Assisted Organic Synthesis (MAOS) has gained an enormous 

attention as an alternative mode of heating to perform a wide variety 

of chemical process. Microwave heating, in contrast to conventional 

heating, enhances the rate of many reactions, improves product yields 

and induces changes in the reaction selectivity [3]. However, up to 

date, there is not a standard method for the on-line monitoring of how 

a reaction proceeds and the optimization of the microwave reaction 

conditions is a matter of trial and error. 

A microfluidic NMR chip with an integrated planar microcoil is 

designed to be coupled to a microwave reactor, with the main aim 

of providing how a particular microwave-assisted continuous-flow 

process develops as a function of time, and how and why it may differ 

under various reaction conditions. A commercial microwave reactor is 

customized, fabricating a microwave flow-cell with a reaction volume 

in the microliter range. The on-line monitoring and optimization of a 

cycloaddition reaction has been performed, with smaller amounts of 

reagents, consuming less energy and in shorter time than when using 

conventional heating, as well as enabling the detection of a very small 

amount of reaction products. 

[1] H. W. Spiess, Angew. Chem., Int. Ed. 2008, 47, 639. 

[2] M.V.Gómez, D.N. Reinhoudt, A.H. Velders, Small, 2008, 4, 1293. 

[3] A. de la Hoz, A. Díaz-Ortiz, A. Moreno. Chem. Soc. Rev., 2005, 34, 

164. 

Fr02 

Nearly 10 6 -fold enhancements in intermolecular 1 H doublequantum 

NMR experiments by nuclear hyperpolarization 

Eliav, Uzi 1 ; Mishkovsky, Mor 2 ; Frydman, Lucio 2 ; Navon, Gil 1 

1 Tel Aviv University, School of Chemistry, Tel Aviv, Israel; 2 Weizmann 

Institute, Chemical Physics Department, Rehovot, Israel 

Intermolecular Multiple-Quantum Coherences (iMQCs) provide a new 

type of contrast in MRI as a result of (a) their dependence on the 

intermolecular distance and (b) the non-linear dependence of the signal 

on the size of the magnetization. We have previously shown that iDQC 

gives enhanced effects of magnetization transfer (MTC) [1], chemical 

exchange saturation transfer (CEST) and nuclear Overhauser effect 

(NOE) [2], in agreement with the theoretical M 2 dependence of the 

signal. A major drawback of iMQC is their inherent low sensitivity. A 

recent study demonstrated that ex situ dynamic nuclear polarization 

(DNP) could assist in overcoming sensitivity problems for iDQC-based 

experiments on 13 C nuclei [3]. In the present work we show that 

although the DNP procedure enhances single-quantum 1 H signals only 

by about 600, which is significantly less than in optimized low-γ liquidstate 

counterparts, the nonlinear dependence of iDQC-derived signals 

on polarization can yield very large enhancements of the 1 H iDQC 

approaching 10 6 . This enhancement allowed us to perform single-scan 

1 H 2D iDQC imaging experiments with SNR comparable to the single 

quantum images but with a different contrast. The short times required 

to obtain 1 H hyperpolarization can be exploited for repetitive injections. 

This in turn opens up interesting possibilities of their own, both in terms 

of imaging applications as well as within spectroscopic settings. 

References: 

[1] U. Eliav and G. Navon, Enhancement of magnetization transfer 

effects by inter-molecular multiple quantum filtered NMR, J. Magn. 

Reson. 190 (2008) 149-153 

[2] W. Ling, U. Eliav, G. Navon, and A. Jerschow, Chemical Exchange 

Saturation Transfer by Intermolecular Double Quantum Coherence, J. 

Magn. Reson. 194 (2008) 29-32. 

[3] E. R. Jenista, R. T. Branca and W. S. Warren, Hyperpolarized carboncarbon 

intermolecular multiple quantum coherences, J. Magn. Reson. 

196 (2009) 74-77. 

Fr03 

Measurement of direct dipolar 1 H- 1 H contacts in magic angle 

spinning solid-state NMR 

Verel, Rene; Schanda, Paul; Huber, Matthias; Manolikas, Theofanis; 

Meier, Beat H. 

ETH Zürich, Physical Chemistry, Zürich, Switzerland 

An experimental method is introduced in which direct dipolar contacts 

between protons are observed. The method employs high Magic Angle 

Spinning rates (up to 55 kHz) at high magnetic fields (19.8 T) and active 

recoupling of the dipolar coupling. To test and characterize the method 

a three dimensional experiment on partially deuterated, microcrystalline 

Ubiquitin was acquired. In this fully resolved and assigned spectrum, 

all of the approximately 40 observed recoupled dipolar contacts could 

be assigned unambiguously. Due to the special nature of the dipolar 

recoupling and the sample preparation, all correlation peaks were 

identified as direct through space dipolar contacts and relayed contacts 

could be excluded. Comparison of the experimentally observed peaks 

with the structure known from X-ray crystallography indicates that all 

contacts correspond to a 1 H- 1 H distance of 6 Å or less. This upper limit 

compares well with information from numerical simulations. Finally, 

many of the correlations are structurally non-trivial and are expected to 

be relevant within the framework of protein structure determination with 

solid-state NMR methods. 


Fr04 

The structure of outer membrane protein A of Mycobacterium 

Tuberculosis 

Teriete, Peter 1 ; Kolodzik, Adrian 1 ; Niederweis, Michael 2 ; Marassi, 

Francesca M. 1 

1 Burnham Institute for Medical Research, La Jolla, CA, United States; 

2 University of Alabama at Birmingham, Department of Microbiology, 

Birmingham, AL, United States 

Approximately 2 million people die from tuberculosis on an annual 

basis and an increasing number of emerging strains of Mycobacterium 

tuberculosis (MTb) show resistance against standard therapeutic 

approaches. Outer membrane proteins in this pathogen are considered 

prime drug targets for new forms of treatment. OmpATb, a newly 

discovered putative outer membrane protein of MTb, is comprised 

of 326 amino acids and was named based on C-terminal sequence 

homology to the porin OmpA of Escherichia coli. 

Tryptic digest studies have identified the central part of OmpATb from 

residue 73 to 220 as the minimal part that is still able to form pores 

and integrate into membranes. Two truncated, His-tagged and isotopelabeled 

forms of OmpATb were successfully expressed in E.coli C41 

cells and purified by affinity chromatography. HSQC spectra show these 

constructs (OmpATb 73-220 and OmpATb 73-326 ) to be stable and folded. The 

HSQC spectrum of OmpATb 73-220 forms an almost perfect subset of the 

HSQC spectrum of OmpATb73-326, indicating that OmpATb 73-220 folds 

independently of the C-terminal part of the protein. 

The solution structure of OmpATb 73-220 has been determined using 

standard NMR methodologies, i.e. heteronuclear edited NOE 

experiments and RDC data in a range of media. Heteronuclear NOE 

and D2O-exchange experiments gave further insight into the dynamics 

indicating a rigid and stable conformation. No comparable arrangement 

of secondary structure elements has been identified amongst currently 

deposited PDBs, suggesting that a novel fold is present in this protein. 

The final structure shows a multi-stranded mixed β-sheet facing 

numerous α-helices in a parallel orientation. A first analysis of the 

functional implication of this structure is presented in this report. 

Biomolecules 


Bi01 

NMR studies of Golgi membrane recognition and proteins in 

solubilizing nanoparticles 

Lenoir, Marc 1 ; Knowles, Timothy J 1 ; Coskun, Ünal 2 ; Buschhorn, Sabine 

B. 2 ; James, Jonathan 1 ; Finka, Rachael 2 ; Smith, Corinne 3 ; Lin, Yu-Pin 2 ; 

Dafforn, Tim 2 ; Simons, Kai 2 ; Overduin, Michael 4 

1 University of Birmingham, Edgbaston, Birmingham, United Kingdom; 

2 Max Planck Institute for Molecular Cell Biology and Genetics, Dresden, 

Germany; 3 University of Warwick, Coventry, United Kingdom; 4 University 

of Birmingham, Edgbaston, Birmingham, United Kingdom 

Membrane proteins are both the most valuable and technically 

challenging targets for drug discovery. Finding a gentle solution 

that preserves their structure and activity yet is robust enough for 

experimental interrogation has represented a major difficulty. A new 

nanoparticle system has been developed that solubilises helical and 

barrel membrane proteins using an unusual amphipathic polymer. 

Proteins maintain their folded structure and binding and enzymatic 

activities in the polymer-lipid nanoparticles based on NMR and 

biophysical assays. The structural mechanisms underlying Golgi 

membrane recognition remain unclear, although the targeting domains 

and role of lipid ligands have been identified. The solution structure 

of the PH domain of the four-phosphate-adaptor protein has been 

determined, revealing an exposed hydrophobic wedge that penetrates 

into mixed micelles designed to mimic the Golgi membrane. Specific 

recognition of PtdIns(4)P and nonspecific engagement of neighbouring 

phospholipid molecules are mediated through a novel binding mode 

and an unprecedented burial of hydrophobic bulk. Deep insertion occurs 

even in the absence of ligand, with a perpendicular orientation of the 

protein on the micelle based on reduced solvation and paramagnetic 

relaxation enhancement of embedded backbone and side chain 

signals. The conservation of key binding features in related PH domains 

indicates that hydrophobic insertion wedge is a dominant feature of 

Golgi recognition, and could deform the bilayer as its lipid components 

are recognized, extracted and trafficked to the cell surface. 

Bi02 

Structure of OmpX in membrane bilayer using NMR 

Lee, Donghan 1 ; Walter, Korvin F.A. 1 ; Brückner, Ann-Kathrin 1 ; Hilty, 

Christian 2 ; Becker, Stefan 1 ; Griesinger, Christian 1 

1 MPI for Biophysical Chemistry, Goettingen, Germany; 2 Texas A&M 

University, Department of Chemsitry, College Station, United States 

Membrane proteins are involved in numerous vital biological processes. 

In order to understand their functions, structures are crucial. However, 

structure determination of membrane proteins is still a frontier in the 

biophysical sciences. In the last few years, solution NMR has been used 

to determine structures of membrane proteins in micelles. Even though 

invaluable information has been obtained, micelles are known to be 

far from ideal in mimicking biological membranes due to their extreme 

curvature and the different lateral pressure compared to the bilayers. 

Furthermore, enzymes often loose their activity in micelles. Recently, we 

have shown that small bicelles contain bilayer part and that membrane 

protein locates in the bilayer part.1 Thus, small bicelles are an excellent 

alternative to mimic the natural environment for membrane proteins 

that allows embedded membrane proteins to retain their activities. 

Here, we present structure of outermembrane protein X (OmpX) from E. 

coli in small bicelles using high-resolution solution NMR spectroscopy. 

Striking differences between structures in bicelles and in micelles were 

observed in area between membrane and aqua environment. 

ABSTRACTS ORAL


1. Lee, D., Walter, K.F.A., Brückner, A.-K., Hilty, C., Becker, S., Griesinger, 

C., Bilayer in small bicelles revealed by lipid-protein interactions using 

NMR spectroscopy. J. Am. Chem. Soc. 2008, 130, 13822-13823. 

Bi03 

Identification of structure conserving motions in the protein 

ubiquitin 

Fenwick, Robert 1 ; Richter, Barbara 2 ; Lakomek, Nils 3 ; Griesinger, 

Christian 3 ; Vendruscolo, Michele 2 ; Salvatella, Xavier 4 

1 IRB Barcelona, Barcelona, Spain; 2 University of Cambridge, Department 

of Chemistry, Cambridge, United Kingdom; 3 Max Planck Institute, 

Biophysical Chemistry, Goettingen, Germany; 4 IRB Barcelona, ICREA, 

Barcelona, Spain 

It has been suggested that motions in proteins could be correlated 

allowing the minimisation of the net changes to the energetic 

contributions that stabilize their native structure (1). Evidence for 

motions in proteins is mounting, however, due to the challenges in 

directly determining time-resolved coordinates from experiments, the 

detection of correlations between the motions of residues distant in 

sequence has remained an elusive goal. Recently the combination of 

NMR with molecular dynamics has been used to characterize protein 

dynamics at atomic resolution (2) and here we use this approach to 

probe the dynamics of the protein ubiquitin. By using the large amount 

of residual dipolar coupling data collected on this protein (3) we have 

generated a large number of conformations collectively consistent with 

experiment and validated the resulting ensemble using independent 

NMR measurements that average on the same time scale. In the 

resulting ensemble we observe the predicted patterns of movement 

that are conserved between neighbouring residues, which are a 

consequence of the peptide geometry. In addition, we now observe 

motions that are a result of the local peptide geometry interacting with 

the hydrogen-bonding network. The motions in beta strands manifest 

in a specific sequence of structural changes and we propose a model 

by which the motion is propagated between the beta strands. Thus, 

the analysis of the ensemble reveals that the backbone motions of 

residues distant in sequence, but connected by hydrogen bonds, not 

only conserve the structure of ubiquitin but may also provide a pathway 

for the transfer of structural and dynamical information across protein, 

demonstrating the underlying mechanisms of binding, allostery and 

folding cooperativity. 

(1) McCammon, Gelin, Karplus, Nature 267, 585 (1977) 

(2) Clore & Schwieters, JACS 126, 2923 (2004); Clore & Schwieters, 

Biochemistry 43, 10678 (2004); Lindorff-Larsen et al., Nature 433, 128 

(2005); Richter et al., JBNMR 34, 117 (2007); Lange et al., Science 

320, 1471 (2008) 

(3) Lakomek et al., JBNMR 34, 101 (2006); Lange et al., Science 320, 

1471 (2008) 

20 

Bi04 

Expression and isotope labeling of G-protein coupled receptors 

in E. coli 

Petrovskaya, Lada 1 ; Bocharova, Olga 1 ; Lyukmanova, Katya 1 ; Shenkarev, 

Zakhar 1 ; Chupin, Vladimir 1 ; Blommers, Marcel 2 ; Arseniev, Alexander 1 

1 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, 

Russian Federation; 2 Novartis Institutes for BioMedical Research, 

Structural Biology Platform, Basel, Switzerland 

G-protein-coupled receptors (GPCRs) belong to a family that consists 

of about 800 trans-membrane proteins of highly pharmaceutical 

relevance. More than half of the drugs that are currently on the market 

target GPCRs. Structure-based drug discovery has been successfully 

applied to soluble proteins, but cannot yet be applied to GPCRs. 

During the last decade, there have been large efforts to develop the 

methodology to study these proteins by X-ray crystallography. These 

activities recently have resulted in the structure determination of a 

few GPCRs. Although this work enhances our knowledge in this field, 

application of the technology in drug discovery is still far away to be 

realized. Apart from X-ray crystallography, that delivers a static picture 

of the structure, NMR spectroscopy would provide in addition knowledge 

of the dynamics of this intrinsically very flexible molecule. Furthermore, 

availability of GPCR samples suitable for NMR, would allow the study of 

interactions with ligands. We have developed the technology to express 

and purify several milligrams of GPCR in E. coli. Critical steps in this 

technology are the use of expression tags that enhance membraneprotein 

expression, auto-induction and slow expression at reduced 

temperatures, the prohibition of disulfide bond driven oligomerization 

by the choice of mutants in which non-essential cysteines are replaced, 

as well as rigorous treatment with reducing agents during refolding and 

purification. We were successful in the expression of various GPCRs 

using these protocols and present here TROSY spectra of the β2adrenergic 

receptor (β2AR) and the melanocortin-2 receptor (MC2R). 

Clear shifts of resonances in the spectra of these 15 N, 2 H-labeled 

proteins in micelles are observed after addition of the receptor-specific 

ligands alprenolol for β2AR and adrenocorticotropic hormone for MC2R, 

thereby validating the quality of GPCR preparation and the feasibility to 

study the interaction between ligands and GPCRs by NMR. 


Computational 

Co01 

NMR spectroscopy beyond Nyquist limitation by random 

sampling and Fourier transform processing 

Kozminski, Wiktor 

University of Warsaw, Department of Chemistry, Warsaw, Poland 

Nuclear Magnetic Resonance is nowadays one of the most efficient 

spectroscopic techniques, providing insight into molecular structure and 

dynamics. The observed frequencies of oscillatory signal are sensitive 

indicators of electron surroundings of nuclei and their, often very subtle, 

changes caused by inter- and intramolecular interactions. However, 

especially in studies of biomolecules, complexity of NMR spectra often 

causes difficulties in their interpretation. This problem may be solved by 

acquisition of multidimensional spectra which increases resolution and 

enables identification of nuclei connected by mutual interactions. 

The conventional approach to recording multidimensional NMR 

experiments is limited by the need for fulfilling of the Nyquist Theorem 

to avoid aliasing. It determines the sampling rate to be twice higher than 

the highest frequency expected in the signal. In consequence, this is an 

implicit limit for the maximum evolution time and therefore obtainable 

resolution. Because of this, for spectra of higher dimensionality, it is 

practically impossible to reach the relaxation limits in indirectly sampled 

dimensions. The problem of effective acquisition of multidimensional 

NMR spectra is getting relatively more important at high fields, where 

the required Nyquist rate increases together with spectral width. 

Random sampling of evolution time space and Multidimensional Fourier 

Transform, enables to obtain, without aliasing, spectra at a very small 

fraction of number of data points required conventionally. Moreover, the 

relative level of artifacts caused by random sampling does not depend 

on this fraction. Some new applications of high dimensionality (3-6D) 

will be shown where the maximum evolution times are limited only by 

transverse relaxation rates. 

K. Kazimierczuk, W. Kozminski, I. Zhukov, J. Magn. Reson. 179, 323- 

328 (2006). 

M. Misiak, W. Kozminski, Magn. Res Chem., 45, 171-174 (2007) 

K. Kazimierczuk, A. Zawadzka, W. Kozminski, J. Magn. Reson. 192, 123- 

130 (2008) 

K. Kazimierczuk, A. Zawadzka, W. Kozminski, I. Zhukov, J. Am. Chem. 

Soc., 130, 5404-5405 (2008) 

K. Kazimierczuk, A. Zawadzka, W. Kozminski, J. Magn. Reson., 197, 

219-228 (2009) 

Co02 

Ensemble calculations for non-structured proteins from PRE 

and RDC data: a case study of 8 M urea unfolded ubiquitin at 

pH 2.5 

Huang, Jie-rong; Grzesiek, Stephan 

Biozentrum, University of Basel, Basel, Switzerland 

The experimental analysis of the unfolded state of a protein provides 

valuable information to guide us to understand of the protein 

folding problem. We have used ubiquitin as a model system for the 

characterization of the unfolded state. A sizeable population of nativelike 

structure within the first β-hairpin of ubiquitin was identified 

under strong denaturing conditions (pH 2.5, 8 M urea) by our previous 

chemical shift, residual dipolar coupling (RDC) and H-bond coupling 

studies (Meier et al. JACS 2007, 129, 754-755). 

A total of 419 RDCs (9 different types) have been obtained from 

steric alignment in strained polyacrylamide gels. To obtain long-range 

information paramagnetic relaxation enhancement (PRE) data (total 

253) were also acquired by using MTSL coupled to eight cysteinemutants 

of ubiquitin. 

In order to get a quantitative description of the unfolded state from 

the experimental data we have developed new modules for both steric 

alignment RDC and PRE ensemble calculations in XPLOR-NIH. The RDC 

module calculates the alignment tensor for each ensemble member 

from its structure at every time-step, and then averages the calculated 

RDCs over the entire ensemble. The potential energy is derived 

from the difference between the ensemble averaged RDCs and the 

experimental data and minimized. Similarly an ensemble PRE module 

uses differences between calculated and observed enhancement of 

transverse relaxation rates for a potential energy term. Both RDC and 

PRE restraints were used simultaneously in the ensemble structure 

calculations. The results for ubiquitin in 8 M urea at pH 2.5 show 

statistically significant conformational propensities with both native- and 

non-native contacts. In particular, a 10-20 % native-like population of 

the first β-hairpin is found, which is consistent with the earlier chemical 

shift and H-bond coupling data. The new modules are generally 

applicable for the characterization of non-structured protein ensembles. 


Co03 

A practical toolbox for NMR solution structure determination in 

structural proteomics 

Gutmanas, Aleksandras 1 ; Lemak, Alexander 1 ; Fares, Christophe 1 ; Wu, 

Bin 1 ; Yee, Adelinda 1 ; Sunnerhagen, Maria 2 ; Arrowsmith, Cheryl 3 

1 University Health Network, Cancer Genomics Proteomics, Toronto, 

Canada; 2 Linköping University, Physics Chemistry and Biology, 

Linköping, Sweden; 3 University of Toronto, Medical Biophysics, Toronto, 

Canada 

The last decade witnessed the steady development of structural 

proteomics initiatives from merely reaching for “low-hanging fruit” to 

solving the structures of ever more challenging proteins. NMR has 

played an important role both as an independent approach to structural 

proteomics and as a salvage pathway for X-ray targets that fail to 

crystallize. Efficiency of data collection and analysis without losses in 

quality of final structures are crucial for the method to remain a viable 

option in the field. 

As part of the Northeast Structural Genomics Consortium (NESG), 

Structural Genomics Consortium (SGC) and Ontario Centre for Structural 

Proteomics (OCSP) our group in Toronto aimed to address both of these 

challenges by developing an overall strategy covering the following 

elements: 

- Identification of an optimal “minimal” set of 2D and 3D NMR spectra. 

- Optimized data acquisition and processing of non-uniformly sampled 

(NUS) 3D spectra. 

- Semi-automatic complete resonance assignment and structure 

determination by Applied BACUS (ABACUS), via a probabilistic 

interpretation of NOESY spectra. 

- Validation and/or refinement of structures with high quality backbone 

RDCs obtained combining NUS and BEST acquisition techniques. 

To further facilitate and speed-up the process, 3 home-built graphical 

user interfaces were designed to guide the user through the steps 

of NUS data acquisition and processing, generation and analysis of 

resonance assignments, and structure calculation and refinement. 

Over the past 3 years using either selected or all of these elements 

we solved and deposited in the PDB 25 structures of novel NESG, SGC 

and OCSP targets as well as proteins from more traditional hypothesisdriven 

research projects, without compromising the quality of the final 

results as verified by torsion angle and steric clash analysis. 

ABSTRACTS ORAL


Co04 

Fully automated solution NMR structure determination 

and recent advances in protein solid-state NMR structure 

determination 

Herrmann, Torsten 

University Lyon, Eurpean Center for High-Field NMR, Lyon, France 

The talk will present computational tools and protocols for automated 

three-dimensional structure determination of proteins by solution and 

solid state NMR. For solution NMR, the unique UNIO protocol for fully 

automated structure determination of proteins will be presented. UNIO 

NMR data analysis interconnects the MATCH algorithm for backbone 

assignment (1), the ASCAN algorithm for side-chain assignment (2) and 

the CANDID algorithm for NOE assignment (3). The ATNOS algorithm 

for robust signal identification in NMR spectra is employed to grant 

each of the three expert-systems direct access to the raw NMR data 

(4), thus enabling integrated and consistent data evaluation. The UNIO 

data analysis protocol requires only a minimal set of 3-6 NMR spectra, 

namely APSY and NOESY, and features high computational efficiency. 

The experience gained so far shows that the UNIO approach leads to 

objective and consistent NMR data interpretation The application range 

and limitations of the fully automated UNIO protocol will be discussed. 

For solid state NMR, a modified version of the ATNOS/CANDID algorithm 

for structural interpretation of solid-state NMR correlation experiments 

will be presented. On the basis of three proteins, the present state of 

automated solid state NMR data analysis using ATNOS/CANDID will be 

discussed. 

(1) Volk, J.; Herrmann, T.; Wüthrich, K. J. Biomol.NMR. 2008, 41, 127- 

138. 

(2) Fiorito, F.; Damberger, F.F.; Herrmann, T.; Wüthrich, K. J. Biomol. 

NMR 2008, 42, 23-33. 

(3) Herrmann, T.; Güntert, P.; Wüthrich, K. J. Mol. Biol. 2002, 319, 209- 

227. 

(4) Herrmann, T.; Güntert, P.; Wüthrich, K. J. Biomol. NMR 2002, 24, 

171-189. 

22 

Enhanced NMR 

En01 

DNP-NMR: Prospects and challenges 

Lerche, Mathilde H 

Imagnia AB, Malmö, Sweden 

The DNP-NMR technique has been given a renewed attention since 

a dissolution protocol for retaining highly polarized spins in liquid 

state was published in 2003. Using this dissolution protocol several 

biochemical applications has been revisited. Much attention has been 

paid to the substrate 1- 13 C-pyruvate as a potential metabolic marker in 

a range of different diseases. The high ambition of reaching a clinical 

application using this technique has, to some extent, taken focus from 

the new tool for solution NMR. DNP-NMR is a useful sensitivity add-on 

tool for several established NMR applications. In addition the technique 

is important in pre-clinical MR where several 13 C-labelled substrates 

have been shown to provide organ and disease specific metabolic 

information. This talk will focus on the prospects for the DNP-NMR 

technique in different biological application areas and provide an 

understanding of the range of applicable compounds. The challenges 

involved in the development of different biochemical and biological 

application areas for this technique are also dealt with to provide an 

expectation matched understanding of the new prospects for DNP-NMR. 

En02 

Overhauser polarization of nitroxide radicals in aqueous 

solutions at high magnetic fields 

Prandolini, M. J.; Gafurov, M.; Denysenkov, V. P.; Sezer, Deniz; 

Endeward, B.; Prisner, T. F. 

Goethe-University Frankfurt, Institute of Physical and Theoretical 

Chemistry, Frankfurt am Main, Germany 

The early development of liquid-state dynamic nuclear polarization (DNP) 

was performed at low magnetic fields, where the Overhauser Effect (OE) 

was found to be most effective. The progress towards high-field DNP 

of large biomolecules in aqueous solutions has been restricted for two 

reasons. Firstly, in the 1960’s it was predicted that the DNP enhancement 

of the OE at higher magnetic fields (over 3 T) rapidly decreases [1]. The 

second reason was the problem of developing microwave components 

and efficient double resonators at higher microwave frequencies. Thus, 

with the development of high-field liquid-state NMR spectrometers, DNP 

was not considered a viable option. In this work we address the first 

problem experimentally comparing actual measured DNP water-proton 

enhancements and power curves together with EPR spectra of various 

isotopic forms of the nitroxide TEMPOL (14N, 15N/ 1H, 2H) at 9.2 T in 

aqueous solutions. 

Liquid water was polarized in-situ at high magnetic fields using a doubleresonance 

structure, which allows simultaneous excitation of the NMR and 

EPR transitions. A custom-made microwave bridge was integrated into a 

standard Bruker 400 MHz NMR spectrometer equipped with a wide-bore 

magnet [2, 3]. We compare the enhancements for both 15N-TEMPOL and 

14N-TEMPOL in water at 318 K. The power curves for the same systems, 

which is the dependence of the inverse enhancement on the inverse MW 

power, predicts a maximum extrapolated DNP enhancement at saturating 

microwave power of between -12 to -20. These results are compared to 

predictions of the DNP coupling factor from nuclear relaxation data [4]. 

[1] K.H. Hausser and D. Stehlik, Adv. Magn. Res. 3, (1968) 79. 

[2] V.P. Denysenkov, M.J. Prandolini, A. Krahn, M. Gafurov, B. Endeward, 

T.F. Prisner, Appl. Magn. Reson. 34, (2008) 289. 

[3] M.J. Prandolini, V.P. Denysenkov, M. Gafurov, B. Endeward, T.F. Prisner, 

submitted 


En03 

Long-lived states to sustain magnetization enhanced by 

dynamic nuclear polarization 

Vasos, Paul R 1 ; Sarkar, Riddhiman 1 ; Ahuja, Puneet 1 ; Comment, Arnaud 2 ; 

Janin, Sami 2 ; Ansermet, Jean-Philippe 2 ; van der Klink, Jacques 2 ; Konter, 

James 3 ; Hautle, Patrick 3 ; van den Brandt, Ben 3 ; Bodenhausen, Geoffrey 4 

1 Ecole Polytechnique Fédérale de Lausanne, ISIC, Lausanne, 

Switzerland; 2 Ecole Polytechnique Fédérale de Lausanne, IPMC, 

Lausanne, Switzerland; 3 Paul Scherrer Institute, Villigen, Switzerland; 

4 Ecole Normale Supérieure, Paris, France 

Major breakthroughs have recently been reported in surpassing two 

main drawbacks of nuclear magnetic resonance spectroscopy: the lack 

of sensitivity and the limited memory of spin states. We have designed 

a combination of dissolution-Dynamic Nuclear Polarization (DNP), as 

originally proposed by Ardenkjaer-Larsen and al., and excitation of 

Long-Lived States (LLS) of nuclear spins. Nuclear magnetization is 

enhanced by up to four orders of magnitude in a polarizing magnet prior 

to transfer of the sample to a high-resolution magnet for detection. In 

the high-resolution magnet, long-lived spin states (LLS) are created and 

sustained by radio-frequency (rf) irradiation to preserve the polarization. 

Enhanced polarization has been maintained in the form of LLS on the 

sensitive proton spins in an Ala-Gly dipeptide during time intervals 

approximately an order of magnitude longer than their spin-lattice 

relaxation time constant T 1 . This opens the way for following slow 

chemical reactions or flow by NMR or MRI. It is promising that LLS of 

the same type can be excited and sustained in amino acids belonging 

to mobile parts of proteins, as relaxation time constants T LLS six times 

longer than the spin-lattice time constants T 1 have been measured for 

residues in the C-terminus of Ubiquitin. 

En04 

Reversible interactions with para-Hydrogen enhance NMR 

sensitivity by polarization transfer 

Duckett, Simon 1 ; Green, Gary 2 ; Cowley, Michael 1 ; Atkinson, Kevin 1 ; 

Ralph, Adams 1 ; Williamson, David 1 

1 University of York, Chemistry, York, United Kingdom; 2 University of York, 

Psychology, York, United Kingdom 

The sensitivity of both nuclear magnetic resonance spectroscopy and 

magnetic resonance imaging is very low because the detected signal 

strength depends on the small population difference between spin 

states even in high magnetic fields. Hyperpolarization methods can be 

used to increase this difference and thereby enhance signal strength. 

This has been achieved previously by incorporating the molecular spin 

singlet para-hydrogen into hydrogenation reaction products. We show 

here that a metal complex can facilitate the reversible interaction of 

para-hydrogen with a suitable organic substrate such that greater than 

5,000-fold increases in proton, with substantial increases in carbon, 

and nitrogen signal strengths, being seen for the substrate without its 

hydrogenation. These signals are then employed in high resolution NMR 

and MRI data collections. 

Some of these results have been communicated (1). 

(1) Paul I. P. Elliott, Simon B. Duckett, Gary G. R. Green, Iman G. Khazal, 

Joaquín López-Serrano and David C. Williamson. Science 2009, 323, 

1709-1711. 

EPR 


Ep01 

Distances and orientations on the nanometer scale from pulsed 

EPR experiments at high fields 

Bennati, Marina 

Max Planck Institute of Biophysical Chemistry, Göttingen, Germany 

Pulsed EPR techniques, in particular the two-frequency DEER or 

PELDOR method, have been established over the past years as a 

valuable tool to obtain distances and their distributions between 

paramagnetic centers in biological systems. When anisotropic 

interactions are resolved in the EPR spectrum at a particular observing 

frequency, these experiments deliver information not only about the 

distance but also about the relative orientation of the paramagnetic 

centres involved. In studies with organic radical probes, the anisotropy 

of the g-tensor can be used for the latter purpose and is best 

resolved in EPR spectroscopy at high-frequencies ( > 90 GHz). In 

the two-frequency experiment, the three-dimensional structure of 

such a paramagnetic spin pair (defined by their distance and relative 

orientation) is encoded in a complex pattern of dipolar modulation 

traces generated by the inherent orientation selection of both pump and 

probe pulses [1-3]. We will present different examples and peculiarities 

of these correlation patterns for endogenous radical pairs in enzymes as 

well as for rigid labels in proteins and illustrate our analysis procedure. 

The experimental design and the technical issues related with the 

performance of these experiments at high EPR frequencies will be 

also discussed in detail. Since the detection range spans distances up 

to several nanometers, the method has a great application potential 

for probing structure and orientations in large protein complexes not 

amenable to X-ray crystallography or NMR. 

[1] V. Denysenkov, T. Prisner, J. Stubbe, M. Bennati, Proc. Natl. Acad. 

Sci. (2006), 103, p. 13389. 

[2] Denysenkov, V., Biglino, D., Lubitz, W., Prisner, T., Bennati, M. Angew. 

Chem. Int. Ed. (2008), 47, p. 1224. 

[3] G. Sicoli, T. Argirevic, J. Stubbe, I. Tkach, M. Bennati, Appl. Magn. 

Reson. (Special Issue) (2009), submitted. 

Ep02 

The solvation of small disulfonate anions in water/methanol 

mixtures characterized by high-field pulse ENDOR 

Hinderberger, Dariush; Heller, Jeannine 

Max Planck Institute for Polymer Research, Mainz, Germany 

The microscopic structure of aqueous solutions has challenged scientist 

for decades. The solvent properties of water, both to solve ionic and 

polar substances, are essential for chemical reactions as well as 

biophysical and biochemical processes. In recent years, much effort has 

been directed into the study of preferential solvation phenomena of ions 

in mixed solvents. 

In this presentation, orientation selective Mims-type electron nuclear 

double resonance (ENDOR) spectroscopy was used to study the 

solvation of Fremy’s Salt (potassium peroxylamine disulfonate, 

K2[ON(SO3)2]) in water/methanol mixtures at 94 GHz (W-Band) in 

glassy solution. We use a simple analysis of the high-field ENDOR 

spectra of different 2H isotope-labeled mixtures of water and methanol 

and can find a distinction of two different exchangeable protons, 

hydrogen-bonded to the unpaired electron spin-bearing NO-group, 

which can be assigned to H-OH of water and H-OCH3 of methanol. 

Furthermore, we find a preferential arrangement of the methyl group of 

methanol along the molecular z-axis of the probe molecule. 

ABSTRACTS ORAL


Ep03 

Site-directed spin labeling pulse EPR reveals the mechanism of 

transmembrane signal transfer in halobacterial phototaxis 

Steinhoff, Heinz-Jürgen 1 ; Döbber, Meike 1 ; Bordignon, Enrica 1 ; Klare, 

Johann 1 ; Holterhues, Julia 1 ; Martell, Swetlana 2 ; Mennes, Nadine 2 ; Li, 

Lin 2 ; Engelhard, Martin 2 

1 University of Osnabrück, Physics, Osnabrück, Germany; 2 Max-Planck- 

Institute for Molecular Physiology, Dortmund, Germany 

Recent developments including pulse techniques and multi-frequency 

approaches make the combination of EPR spectroscopy, site-directed 

spin labeling (SDSL), and molecular dynamic simulations an attractive 

approach for studying the structure and conformational dynamics of 

membrane protein complexes. Analysis of the spin label side chain 

mobility, its solvent accessibility, the polarity of the spin label microenvironment 

and distances between spin label side chains provide 

information for restraint modeling of membrane protein conformations 

with high temporal and spatial resolution [1]. The presentation first 

reviews our recent results on the application of pulse ELDOR methods 

on membrane bound colicin A, vinculin, and the ABC-importer MalFGK 2 

[2]. We then focus on our multi-frequency EPR and pulse ELDOR 

studies of the halobacterial phototaxis receptor sensory rhodopsin 

(pSRII) in complex with the receptor specific transducer (pHtrII). This 

complex is considered as a general model system for transmembrane 

signal transduction. Inter-spin distances determined from pairs of 

interacting nitroxide spin labels lead to a unique structural model of 

the dimeric complex [3]. Time resolved detection of inter-spin distance 

changes after light activation reveals conformational changes of pSRII 

and uncovers the mechanism of the signal transfer from pSRII to the 

associated transducer pHtrII [4]. Subsequent conformational changes in 

the first HAMP domain of pHtrII are shown to be the language of signal 

transfer [5]. 

References: 

[1] E. Bordignon and H.-J. Steinhoff. In: ESR Spectroscopy in Membrane 

Biophysics. (Hemminga, M.A., and Berliner, L.J., eds). Springer Science 

and Business Media, New York (2007) 129-164; 

[2] M. Grote et al. (2008) Biophys. J. doi: 10.1529/ 

biophysj.108.132456; 

[3] A. A. Wegener, et al. (2001) EMBO J. 20: 5312-9; 

[4] E. Bordignon et al. (2007) Photochem. and Photobiol. 83: 263–272; 

[5] M. A. Döbber et al. (2008) J. Biol. Chem. 283: 28691-28701. 

Ep04 

Resolving the 14 N and 1 H frequencies of the two Cu(II) sites 

in ascorbate oxidase by high field pulse EPR correlation 

spectroscopy 

Potapov, Alexey; Goldfarb, Daniella 

Weizmann Institute, Chemical Physics Dept., Rehovot, Israel 

Electron-nuclear double resonance (ENDOR) and electron-electron 

double resonance (ELDOR)-detected NMR spectroscopic techniques 

are methods for measuring nuclear frequencies in paramagnetic 

systems. The spectra obtained are often congested due to the presence 

of multiple paramagnetic species and multiple nuclei. Therefore 

correlation techniques that can resolve and assign lines are important, 

particularly if they can replace biochemical preparative procedures. 

Such an experiment, THYCOS (triple resonance sublevel correlation 

spectroscopy) has been recently described [1]. It provides links between 

forbidden electron spin transitions (M S = ± 1, M S ≠0) and allowed 

nuclear spin transitions (M I =±1), thus, facilitating the assignment of 

nuclear frequencies to their respective electron spin manifolds and 

paramagnetic centres. It also yields the relative signs of the hyperfine 

couplings of the different nuclei. 

24 

The feasibility and the information content of the method were 

demonstrated on a single crystal of Cu-doped L-histidine and on a 

frozen solution of a Cu-histidine complex[1]. Here we apply THYCOS, 

corroborated with HYSCORE (Hyperfine sublebvel correlation), both 

carried out at a high field (~ 3.5 T, 95 GHz), to the enzyme ascorbate 

oxidase, which consists of two EPR active Cu(II) sites. The type 1 and 

type 2 coppers were discriminated based on correlation of the 14 N 

signals with the large 1 H couplings of the cystein β-protons. The spectra 

showed that the 14 N nuclei with a hypefine couplings of ~20 MHz 

belong to type I whereas those with the ~40 MHz to type II. This result 

shows that this method can be useful for studies of other biomolecules 

containing multiple paramagnetic sites. [1] Alexey Potapov, Boris Epel, 

and Daniella Goldfarb J. Chem. Phys. 128, 052320 (2008) 


Im01 

Imaging 

MRI measurements of fluid flows 

Gladden, Lynn 

University of Cambridge, Department of Chemical Engineering, 

Cambridge, United Kingdom 

Magnetic resonance velocimetry techniques are increasingly being used 

to study flow phenomena in non-medical systems. This presentation will 

be illustrated with examples of the types of measurements that can be 

made and how the resulting information can be used. Measurement of 

fluid flows over the range of hundreds of microns per second to metres 

per second will be shown. 

Amongst the recent case studies that have been undertaken by the 

group, the following will be highlighted to illustrate the diversity of 

measurements that can be made using magnetic resonance techniques. 

For example: (i) crystallisation processes; (ii) bifurcation phenomena 

in laminar and time-dependent flows through a sudden expansion; (iii) 

two-phase flows of gas and solids; and (iv) gas-liquid flows in porous 

media such as catalytic reactors. In the final example, the ability to track 

chemical composition within reactor environments will also be shown. 

The motivation for each case study will be described briefly and it will 

be shown how a variety of different magnetic resonance techniques can 

be used to obtain the required insights into a given problem. 

Finally, consideration will be given as to how we can increase data 

acquisition rates. ‘Ultra-fast’ acquisition times for two-dimensional flow 

maps are typically 20 ms. Recently we have been exploring the use of 

compressed sensing techniques to further decrease data acquisition 

times and the latest results will be presented. This final topic addresses 

a field of research which is of general application in MRI, and has 

already attracted considerable attention in the medical community. 

Im02 

Magnetic Resonance imaging in inhomogeneous fields 

Paquin, Raphael; Pelupessy, Philippe; Bodenhausen, Geoffrey 

Ecole Normale Superieure, Department of Chemistry, Paris, France 

Single scan NMR spectroscopy, introduced by Frydman and co-workers 

few years ago [1], has been extended recently to obtain magnetic 

resonance images. A subsequent study presented the efficiency of 

tailored pulses and gradients to compensate a priori known Zeeman 

field inhomogeneities [2]. We propose a single scan scheme to 

counteract the effect of field inhomogeneities without mapping them 

previously, based on adiabatic pulses [3] and simultaneous application 

of two orthogonal gradients for encoding and decoding. Acquisition of 

2D and 3D images are obtained in inhomogeneous fields characterized 

by a 3000 Hz breadth of water resonance. 

[1] L. Frydman, T. Scherf, A. Lupulescu, The acquisition of multidimensional 

NMR spectra within single scan, Proc. Natl. Acad. Sci. USA, 

99, (2002), 15858-15862. 

[2] A. Tal, L. Frydman, Spatially encoding and the single-scan acquisition 

of high definition MR images in inhomogeneous fields, J. Magn. Res., 

182, (2006), 179-194, 

[3] P. Pelupessy, Adiabatic single scan two-dimensional NMR 

spectroscopy, J. Am. Chem. Soc., 125, (2003), 12345-12350. 


Im03 

Mapping of drug recrystallization in solid dispersions by 1 H and 

19 F NMR and NMR microimaging 

Dahlberg, Carina 1 ; V. Dvinskikh, Sergey 1 ; Schuleit, Michael 2 ; Furó, 

István 1 

1 Royal Institute of Technology, KTH, Physical Chemistry and Industrial 

NMR Centre, Stockholm, Sweden; 2 Novartis Pharma AG, Pharmaceutical 

and Analytical Development, Basel, Switzerland 

Many potentially powerful drugs are poorly soluble in aqueous media. 

One favourable approach for utilizing them is via forming solid 

dispersions with the drug in amorphous form embedded in a carrier 

matrix. Such formulations often enhance bioavailability and dissolution 

rate. However, poor physical stability, evidenced by crystallization, is one 

major disadvantage and barrier to the marketing success. Exploiting the 

full potential of formulations with drugs in their amorphous state require 

that they remain stable in solid state, as well as during dissolution. 

Understanding why certain pharmaceuticals are stable in their 

amorphous form any why others rapidly recrystallize, or require storage 

at special conditions to confer stability, is of paramount importance. 

Chemical and physical instability of pharmaceuticals containing 

amorphous drugs and/or exipients is ascribable, in large part, to their 

higher molecular mobility compared with crystalline solids. This, under 

favourable circumstances, also provides one with the opportunity 

to observe crystallization through the spin relaxation behaviour. 

Hence, we demonstrate an NMR relaxation method to follow the 

recrystallization process in pharmaceutical systems. This method 

allows us to characterize the change of state of drugs and carriers over 

pharmaceutically significant time scales. Unlike bulk methods, NMR 

relaxation imaging has the advantage of providing spatially resolved 

information about recrystallization under relevant processes such as 

tablet dissolution. By mapping the effect of drug properties, drug-carrier 

interactions, carrier stabilization and manufacturing process, the best 

strategy to overcome the recrystallization drawback is revealed for 

certain drugs. 

Im04 

Overview of applications for ultra-low field magnetic resonance 

Sandin, Henrik; Araya, Yonathan; Flynn, Mark; Gomez, John; Hanson, 

Christina; Kraus, Robert; Magnelind, Per; Maskaly, Karlene; Matlashov, 

Andrei; Nath, Pulak; Newman, Shaun; Owens, Tuba; Peters, Mark; 

Savukov, Igor; Schultz, Larry; Urbaitis, Al; Volegov, Petr; Zotev, Vadim; 

Espy, Michelle 

Los Alamos National laboratory, Los Alamos, United States 

Magnetic resonance (MR) techniques have been around for over 50 

years and the trend has been to go to higher and higher field strengths, 

now approaching 7 Tesla. The idea for magnetic resonance in the 

earth’s magnetic field has been proposed, but the very low polarizations 

precluded any practical application. More recently it has been shown 

to be possible and practical to perform “ultra-low field” (ULF) MR 

which works in the regime of µT to mT field strengths. With fields only 

a fraction of the strength of high field MR the expected signals are 

very small. To be able to detect these signals we use Superconducting 

QUantum Interference Devices or SQUIDs. SQUIDs are the most 

sensitive sensor that exists for magnetic sensing and they can detect 

magnetic fields down in the fT range. In the past decade the potential 

areas of applications of ULF MR have grown to include areas as diverse 

as medical MRI and detecting bomb threats at airports. The ULF MR 

approach gives new opportunities to explore scientific areas that 

have not been possible at high fields. With ULF working in the 10-10 4 

Hz Larmor frequency range it has an interesting overlap with many 

biological processes. The technique also uses a pulsed pre-polarizing 

field that has no dependence to the measuring field or gradient 

ABSTRACTS ORAL


strengths. This gives the approach a very flexible setup which allows 

quick and easy changes in the configuration of data acquisition, pulse 

sequences, and orientation of all fields. Other benefits of the technique 

are the ability to image through non-ferrous metal containers and image 

parts of a human body that contain non-ferrous metal. In this talk I will 

describe the motivation and the progress the team has made in many 

different areas of ULF MR. This includes areas of MRI, classification 

and detection of hazardous material at an airport using a ULF MR 

relaxometer, and how weak ( ~10µA) currents can interact and change 

the signal response from spin-populations in water phantoms. 

26 

Metabolomics 

Me01 

NMR based metabolomics in non genotoxic carcinogenicity: 

methods for expanding the coverage of the metabolome 

Waterman, Claire; Rubtsov, Denis; Griffin, Julian 

University of Cambridge, Department of Biochemistry, Cambridge, 

United Kingdom 

Non-genotoxic carcinogens are particularly difficult to identify in the 

safety assessment process of pharmaceuticals and agrochemicals as 

they do not damage DNA directly and have diverse modes of action, 

necessitating long term in vivo studies. We have been applying a 

combined metabolomic, transcriptomic and histological approach to 

study the action of non-genotoxic carcinogens with the intention of 

investigating early stage mechanistic changes that are predictive of 

longer term pathology. This presentation will focus on the use of NMR 

spectroscopy based metabolomics and how we have used a variety of 

approaches to increase the sensitivity and coverage of this approach. 

Firstly, to increase the coverage of the network of metabolism within 

liver tissue data fusion was employed between the NMR spectroscopic 

dataset and the acquired transcriptomics. This approach was 

particularly useful for understanding why metabolites found at the hubs 

of several pathways were perturbed by a given modification. Secondly, 

a novel Bayesian approach to processing FIDs has increased metabolite 

detection within these datasets, as well as produced datasets which 

contain only resonances, and hence do not import noise into the 

subsequent multivariate analysis. Thirdly, high resolution magic angle 

spinning (HRMAS) 1H NMR spectroscopy has been used to profile 

both aqueous and lipophilic metabolites to better understand the 

consequences of a given lesion. 

Me02 

Robust 1 H NMR-based metabolomic responses in fish exposed 

to different sewage effluents in two separate studies 

Samuelsson, Linda 1 ; Björlenius, Berndt 2 ; Förlin, Lars 3 ; Larsson, Joakim 1 

1 University of Gothenburg, Institute for Neuroscience and Physiology, 

Göteborg, Sweden; 2 Stockholm Water Co., Stockholm, Sweden; 

3 University of Gothenburg, Department of Zoology/Zoophysiology, 

Göteborg, Sweden 

Sewage effluents contain complex mixtures of contaminants such as 

metals, flame retardants, surfactants, pharmaceuticals and household 

chemicals. Many of these micropollutants are not removed in our 

sewage treatment plants. Thus, there are concerns about potentially 

adverse effects on aquatic organisms, and the addition of advanced 

treatment steps in existing plants has been suggested. A biological 

evaluation of the effects on organisms exposed to differently treated 

effluents is needed as a complement to chemical analyses, especially in 

cases where new, unknown and potentially harmful compounds may be 

formed during the treatment process. As the biological modes of action 

of new compounds are not known, explorative methods are required. 

1 H NMR-based metabolomics, a method for simultaneous analysis of 

up to 100 endogenous small metabolites in an organism, tissue or 

biofluid, was used in this work. In the first study, rainbow trout were 

exposed in parallel to six differently treated effluents for two weeks 

and blood plasma was analysed by 1D 1 H NMR. Multivariate data 

analysis showed changes in the metabolome (increases in HDL and LDL 

lipids, cholesterol, glucose, phosphatidyl choline, glutamine, alanine 

and decreases in VLDL lipids and glycerolipids) caused by exposure 

to some effluents. This was the basis to postulate a hypothesis that 

certain treatment processes generated compounds affecting the 


metabolic profile of fish. The hypothesis was tested and confirmed in an 

independent study the following year. This allowed an identification of 

treatment processes generating potentially harmful compounds. 

To conclude, 1 H NMR-based metabolomics proved to be an open or 

explorative method suitable for identifying the presence of unknown 

compounds in a mixture with a capacity to affect the metabolite profile 

of exposed organisms. The metabolic responses identified were found 

to be robust between two independent studies. Although we cannot yet 

link the metabolic changes to adverse effects, the results show a great 

promise for metabolomics in risk assessment of complex mixtures, 

where there identity and mode of action of pollutants are unknown. 

Me03 

Real time observation of TCA cycle metabolites by DNP 

Marin-Montesinos, Ildefonso; Lodi, Alessia; Carrigan, John; Saunders, 

Martin; Ludwig, Christian; Gunther, Ulrich 

University of Birmingham, School of Cancer Studies, Birmingham, United 

Kingdom 

Dynamic Nuclear Polarisation (DNP) is used to transfer the high spin 

polarization of unpaired electrons to coupled nuclear spins. Stable 

radicals are added to a solution of a target sample and irradiation 

with microwaves is applied at the EPR lines of the radical. In such 

experiments, enhancements of >10,000 were achieved [1] after rapidly 

warming up polarised samples to approx. 300K, and transfer to a high 

field magnet where spectra are recorded. DNP bears significant new 

possibilities for the application of NMR in metabolomics, in particular for 

flux measurements. 

It is well known since Warburg’s hypothesis in 1924 that cancer affects 

energy metabolism. Recent work used this principle to observe real time 

metabolism of isotopically labelled pyruvate in animals [2], cancer cells 

and tumour tissue [3,4]. In these previous studies, hyperpolarised 13 C 1 - 

pyruvate was either fermented to lactate or transaminated to alanine 

through enzymatic processes in the cell. However, further insights can 

be expected from using alternative pyruvate labelling schemes or other 

labelled metabolic precursors. 

Here we demonstrate, for the first time, that it is possible to observe 

real time generation of TCA cycle metabolites in mitochondria from 

acute myeloid leukemia cell lines using the HyperSense TM polariser from 

Oxford Instruments. Different pyruvate labelling schemes are employed 

in order to maximise the number of metabolites being observed. Then, 

these types of experiments can be used to study TCA cycle dysfunction 

and to understand drug induced responses which cause apoptosis or 

differentiation in cellular models. 

References 

[1] JH Ardenkjær-Larsen, B Fridlund, A Gram, G Hansson, L Hansson, 

MH Lerche, R Servin, M Thaning, and K Golman. Proc Nat Acad Sci USA, 

100, 10158-10163 (2003) 

[2] K. Golman, R. Zandt, and M. Thaning. Proc. Nat. Acad. Sci. USA, 

103, 11270-11275 (2005) 

[3] M. E. Merrit, C. Harrison, C. Storey, F. M. Jeffrey, A.D. Sherry, and C. 

Malloy. Proc. Nat. Acad. Sci. USA, 104, 19773-19777 (2005) 

[4] S. E. Day, M. I. Kettunen, F. A. Gallagher, D. E. Hu, M. Lerche, J. 

Wolber, K. Golman, J. H. Ardenkjaer-Larsen, and K. M. Brindle.Nature 

Medicine, 13, 1382-1387, (2007) 


Me04 

Intramolecular isotope distributions reveal metabolic regulation 

Schleucher, Jürgen 

Umea University, Medical Biochemistry & Biophysics, Umea, Sweden 

Metabolomics or analytical sciences in general measure concentrations 

or activities, for example of metabolites or enzymes. However, the 

regulation of a pathway is often of greatest interest, but there is 

no direct link between concentrations / activities and regulation. 

In contrast, intramolecular distributions of heavy stable isotopes, 

measurable by NMR, are directly sensitive to metabolic regulation. 

Stable heavy isotopes are omnipresent, and variation in their 

abundances turns them into tracers. During the formation of any 

compound, physical and chemical fractionations of heavy isotopes 

(isotope effects) modulate the amount of isotopes that get incorporated 

into the molecule as a whole, and into each distinct intramolecular 

group, so-called isotopomer (e.g. glucose with 13 C at C2). Thus, 

isotopomer abundances are directly linked to chemical reaction 

mechanisms and to the regulation (e.g. rate-limiting steps) of synthesis 

and breakdown. Because NMR, as nondestructive method, eliminates 

the possibility of isotope fractionation during measurement, it is the 

prime tool to measure isotopomer abundances. 

Here we describe NMR methodology to measure D and 13 C isotopomers, 

and present concepts for the interpretation of isotopomer data. We 

demonstrate that i) Alternative synthetic pathways create characteristic 

isotopomer patterns, ii) Within a given synthetic pathway, flux control 

exerted by individual reaction steps can be deduced, iii) When both an 

organism’s environment and its biochemistry cause variation in isotope 

abundance, isotopomer distributions reflect responses of organisms to 

their environment. Isotopomer distributions advance our understanding 

of biological systems in two ways. Firstly, isotopomers directly reflect 

the relative contributions of competing pathways and the regulation of 

individual pathways, without perturbing the organism studied. Secondly, 

information on pathway regulation is stored in metabolites, and can be 

obtained retroactively, which opens exciting possibilities for the study of 

plant-climate interactions on long time scales, of sources and sinks of 

pollutants in the environment, and of changes in metabolism linked to 

diseases. 

ABSTRACTS ORAL


28 

Molecular Interactions 

Mi01 

NMR studies of proteins and RNAs in the regulation of gene 

expression 

Sattler, Michael 

Helmholtz Zentrum München, Neuherberg, Germany; Technische 

Universität München, Garching, Germany 

We are studying mechanisms of protein and RNA recognition in the 

regulation of pre-mRNA splicing and gene silencing by non-coding 

RNAs. 

Splicing of nuclear pre-mRNA, i.e. the removal of non-coding, 

intervening intron sequences, is a key step in the regulation of 

eukaryotic gene expression. It contributes to gene regulation and protein 

diversity by joining of alternative exons. Early spliceosome assembly 

is tightly regulated and involves the recognition of characteristic 

intron RNA sequences. The biogenesis of these pre-spliceosomal 

intermediates is dynamic and involves cooperative binding to the intron 

RNA. Results will be presented on our structural studies of protein-RNA 

and protein-protein interactions that play important roles in spliceosome 

assembly and the regulation of alternative splicing. 

Xist (X-inactivation specific transcript) RNA is a large non-coding RNA 

essential for the initiation of X-chromosome inactivation in female 

mammals. It is expressed from the X-chromosome to be silenced and 

coats it in cis, which coincides with transcriptional shutdown through a 

largely unknown mechanism. We are performing structural studies of 

the molecular mechanism of X-inactivation involving Xist RNA. 

For structural analysis of multi-domain proteins and protein complexes, 

we are exploring strategies to determine the quaternary arrangement of 

multimeric protein complexes by solution NMR methods. We combine 

orientational information derived from residual dipolar couplings 

(RDCs) and (long-range) distance restraints derived from paramagnetic 

relaxation enhancement (PRE) using spin-labeled proteins and/or RNA. 

In addition, we have implemented a novel target function in CNS for 

direct refinement against small angle X-ray and/or neutron scattering 

(SAXS/SANS) data. The RDC, PRE and SAS data can be jointly used for 

structure calculation in ARIA/CNS and be supplement with additional 

information from chemical shift perturbation or biochemical data. 

Mi02 

Phosphorylation and membrane association of the natively 

unfolded c-Src unique domain 

Perez, Yolanda 1 ; Bernadó, Pau 1 ; Gairí, Marga 2 ; Pons, Miquel 1 

1 Institute for Research in Biomedicine, Barcelona, Spain; 2 University of 

Barcelona, Barcelona, Spain 

c-Src is a membrane-bound, non-receptor tyrosine kinase involved 

in numerous signal transduction pathways. Localization to different 

membrane compartments is probably essential for discrimination 

between different pathways. Membrane localization of c-Src involves 

its unfolded N-terminal domain (USrc), including a N-terminal myristoyl 

group and 6 positively charged amino acids, part of the SH4 domain. 

Membrane binding increases with the amount of acidic lipids (1). 

Previous studies suggested that membrane attachment (2) could 

involve additional unspecified regions. The localization of membrane 

interaction regions, their structural significance, their interaction with 

the SH4 (unfolded) and SH3 (folded) domain, and their modulation by 

phosphorylation will be presented. 

We have studied the unfolded N-terminal region of human c-Src (USrc, 

84 aa long) (3) using NMR in presence of bicelles formed by a mixture 

of short- and long-chain phospholipids (4,5). Well resolved spectra were 

observed showing significant lipid dependent chemical shift changes in 

two regions flanked by residues 7-23 and 49-68. 

Four USrc phosphorylation sites (S12, S17, T37, and S74) have 

been described. Phosphorylation in the N-terminal region results in a 

reduction of the affinity to the plasma membrane (6). We have studied 

by NMR the effect of in vitro phosphorylation by PKA (S17) or CDK5/p25 

(T37, S74) on the interactions with acidic bicelles and have investigated 

the structural effects of phosphorylation at these three sites using 

RDCs. 

Our work highlights the use of NMR to study the interaction of unfolded 

domains with membranes. 

1. Murray,D. et al. Biochemistry, 37, 2145, 1998. 

2. Kaplan, J.M., Varmus, H.E. and Bishop, J.M. Mol. Cell. Biol., 10, 

1000, 1990. 

3. Pérez, Y., Gairí, M., Pons, M. and Bernadó. In preparation. 

4. Poget, S.F. and Girvin, M.E. BBA, 1768, 3098, 2007. 

5. Struppe, J., Whiles, J.A. and Vold, R.R. Biophys. J., 78, 281, 2000. 

6. Walker, F., deBlaquiere, J. and Burges, A.W. J. Bio. Chem., 268, 

19552, 1993. 

Mi03 

Characterization of an enzyme reaction by STD NMR 

spectroscopy 

Hackl, Thomas 1 ; Fransson, Iris G. 2 ; Meyer, Bernd 3 

1 University of Hannover, Insitute of Organic Chemistry, Hannover, 

Germany; 2 University of Hamburg, Department of Biochemistry, 

Hamburg, Germany; 3 University of Hamburg, Institute of Organic 

Chemistry, Hamburg, Germany 

Investigation of protein ligand interactions by NMR based methods in 

solution provides insight into the binding process at an atomic level. 

STD NMR (saturation transfer difference) is a ligand based approach, 

which has proven to be applicable to a broad spectrum of proteins. In 

addition to soluble proteins also membrane-bound receptors integrated 

in liposomes or cells can be examined. A few examples studying 

enzymatic systems have been reported. 

Investigation of substrate binding to an enzyme is achieved easily, if 

two substrates are necessary for the reaction. However, in the case of 

a one-substrate enzyme there will be conversion to the product during 

the experiment. A STD NMR experiment will be presented that allows 

the characterization of substrate and product binding epitopes and 

additionally gives access to kinetic data. The experiment was performed 

on germacren D synthase, an enzyme that belongs to the family of 

sesquiterpene synthases. These enzymes catalyze complex multistep 

reactions from a common precursor, farnesyl diphosphate, and are 

capable of generating hundreds of structurally diverse hydrocarbon 

scaffolds. 

A property of terpene synthases is their low turn over number. 

Germacene D synthase has a k cat = 0.02 s -1 [1]. On the other hand 

the enzyme revealed a much higher dissociation rate of the enzyme 

substrate complex k -1 = 1.2 s -1 , assigned by SPR (surface plasmon 

resonance) experiments. These properties, the low k cat and k -1 >>k cat , 

provide excellent conditions to perform STD NMR. A pseudo-2D STD 

pulse sequence was modified, acquiring STD data for short time 

intervals. Each FID of subsequent time intervals is stored in subsequent 

increments of the 2D data file. The off-resonance spectra exhibit 

the reaction progress, which can be analyzed by computer assisted 

methods (progress curve analysis). The STD difference spectra allow 

an insight into the binding process. Additionally, corresponding FIDs of 

subsequent time intervals can be summed up to increase the signal to 

noise in STD spectra retrospectively. 

[1] I. Prosser, I.G. Altug, A.L. Phillips, W.A. König, H.J. Bouwmeester, 

M.H. Beale, ABB, 432, 2004, 136–144. 


Mi04 

Studying protein binding with solid-state NMR: the interaction 

of an antifreeze protein with ice 

Siemer, Ansgar; McDermott, Ann 

Columbia University, Chemistry, New York, United States 

Liquid-state and solid-state NMR were combined to investigate an 

unusual protein-ligand interaction; the binding of an antifreeze protein 

to ice. 

Antifreeze proteins (AFPs) can be found in a variety of different 

organisms such as fish, insects, plants, and bacteria. Binding to ice, 

AFPs lower the freezing point of a given solution below its melting point. 

Our research focuses on one of the best studied AFPs, a type III AFP 

(AFP III) found in arctic fish. Type III AFPs are small, globular, mixed α/β 

proteins of 62-66 amino acids in length. High-resolution X-ray and 

liquid-state NMR structures of a type III AFP together with site-directed 

mutagenesis experiments and molecular dynamics (MD) simulations, 

suggested an ice-binding site for AFP III. This putative ice-binding site is 

formed by a flat surface on the protein. 

Using a chemical-shift perturbation study we compared chemical 

shifts of AFP III in the frozen, solid state with the chemicals shifts of 

AFP III in the unfrozen liquid state and delivered further evidence of the 

ice-binding site of this protein. Comparing AFP III to ubiquitin in frozen 

solution and we found remarkable differences in their 1 H T 1 relaxation 

rates and their chemical-shift perturbation upon freezing, although both 

proteins gave relatively high resolution spectra in frozen solution. 

We will also present our first results towards the direct measurement of 

ice-AFP III interactions and 1 H chemical-shift perturbation data of AFP III. 

Paramagnetic 


Pa01 

Structures and dynamics of weak protein complexes studied 

with paramagnetic NMR tools 

Ubbink, Marcellus; Prudencio, Miguel; Vlasie, Monica; Keizers, Peter; 

Xu, Xingfu; Lindfors, Hanna; Hass, Mathias; Volkov, Alexander; Worrall, 

Jonathan; Overhand, Mark 

Leiden University, Institute of Chemistry, Leiden, Netherlands 

Many protein complexes are weak and short-lived, because their 

biological function requires fast turn-over. Electron transfer and cell 

signalling complexes are among the most transient of complexes, with 

lifetimes as low as 1 ms and dissociation constants in the micromolar 

range. 

We use paramagnetic NMR methods to characterize the structure and 

dynamics of these complexes. A paramagnetic center is engineered 

specifically on one of the two proteins. By using such tags, we 

have recently determined the structures of the 46 kDa complex of 

cytochrome c and cytochrome c peroxidase and the 152 kDa complex 

of nitrite reductase and pseudoazurin in solution. 

Paramagnetic NMR also makes it possible to probe the dynamics within 

protein complexes. The strong and highly distance-dependent nature 

of the paramagnetic effects allows the detection of conformations that 

represent only a small fraction of the total population. Recently, we 

studied the dynamics in various protein complexes. Peptide binding 

to SH2 and SH3 domains using the rigid TOAC spin label shows, 

surprizingly, that binding is less dynamic for SH3 than for SH2 domains. 

Another complex with dynamic nature is the electron transfer complex 

of cytochrome c and adrenodoxin. A comparison of the native complex 

with a crosslinked counterpart shows that in the former extensive 

averaging of all measured NMR parameters occurs, suggesting that 

this complex exists entirely as a dynamic complex, representing a pure 

encounter state . 

In conclusion, paramagnetic NMR experiments show that weak protein 

complexes have a highly dynamic nature in solution, which cannot be 

captured by static structures determined with classical NMR or X-ray 

diffraction. These complexes are best described by an equilibrium 

between a well-defined state and a dynamic encounter state. In extreme 

cases, this equilibrium can be shifted strongly towards the dynamic 

side. 

Keizers et al. J. Am. Chem. Soc. 2008, 130, 14802-14812; 

Volkov et al. Proc. Natl. Acad. Sci. USA 2006, 103, 18945-18950; 

Vlasie et al. J. Mol. Biol. 2008, 375, 1405-1415; 

Lindfors et al. J. Biomol. NMR 2008, 41, 157-167; 

Xu et al. J. Am. Chem. Soc. 2008, 130, 6395-6403 

Pa02 

The motions of domains, as seen by RDCs 

d’Auvergne, Edward; Coudevylle, Nicolas; Dorr, Brent; Rodriguez- 

Castaneda, Fernando; Frank, Benedikt; Fares, Christophe; Griesinger, 

Christian 

MPIBPC, Goettingen, Germany 

Through the selective alignment of individual domains, the time 

averaged, absolute motions of multi-domain macromolecules can 

be probed by the observation of reduced residual dipolar couplings 

(RDCs). This is often achieved by internal, paramagnetic alignment. A 

number of theories have been developed for the interpretation of the 

alignment tensor reduction including the N-state model, the pmax and 

MAP(R) theories, and trajectory based approaches. In the N-state or 

ABSTRACTS ORAL


discrete state models, the individual states or structures can easily 

be shown to be non-representative via basic test models. Yet the 

average properties of the ensemble of N-states match those of the true 

model. Here this conserved information in the N-state model will be 

explored using the geometric interpretation of the ball-and-socket or 

spherical joint mechanical model. Using this model, the average domain 

orientation and the amplitudes of certain components of the motions, as 

represented by cone angles or order parameters, will be extracted from 

the N-state model fitted to the RDCs. The mechanical joint model can 

be used universally to interpret domain motions in molecular systems, 

and will be applied to calmodulin complexed to target peptides. 

Pa03 

Solid-state NMR of microcrystalline paramagnetic Cu(II) 

protein: relaxation-optimized sequences, ultra-fast MAS and 

structural constraints 

Laage, Ségolène 1 ; Pierattelli, Roberta 2 ; Felli, Isabella 2 ; Bertini, Ivano 2 ; 

Sachleben, Joseph R. 3 ; Steuernagel, Stefan 4 ; Lesage, Anne 1 ; Emsley, 

Lyndon 1 ; Pintacuda, Guido 1 

1 Université de Lyon, CNRS/ ENS Lyon/ UCB-Lyon 1, Centre RMN à 

Très Hauts Champs, Lyon, France; 2 CERM, University of Florence, 

Department of Chemistry and Magnetic Resonance Cen, Sesto 

Fiorentino, Italy; 3 Otterbein College, Columbus, OH, United States; 

4 Bruker Biospin, Karlsruhe, Germany 

We present our recent advances in the structural investigation by 

solid-state magic angle spinning (MAS) NMR of a microcrystalline 

paramagnetic protein, the human superoxide dismutase (SOD), 

a dimeric Cu(II) enzyme of 32 kDa. In this system, sensitivity and 

resolution represent a challenge due to the large size of the molecule 

and the short relaxation times, consequence of the hyperfine interaction 

between the nuclei and the unpaired electrons of Cu(II).[1] 

First, to gain access to crowded spectral regions, we introduce 

relaxation-optimized methods for 13C-13C spin-state selection, which 

remove the broadening due to the 13C-13C J couplings and lead to 

a considerable enhancement in both resolution and sensitivity in 2D 

experiments.[2] 

Second, we explore the impact of so-called ultrafast (>60 kHz) MAS 

in the characterization of biomolecular solids, notably containing 

paramagnetic centers. We discuss a set of experiments relying on 

low-power rf field irradiation (low-power decoupling schemes, bandselective 

Hartmann−Hahn cross-polarization between the whole proton 

bath and carbons whose resonances are close to the 13C-transmitter 

offset). Combined with the short 1H T1s, these schemes allow the 

repetition rate of the experiments to be significantly shortened.[3,4] 

Finally, we show how evaluation of some of the paramagnetic effects 

(relaxation rates and anisotropic shifts) can provide a precious set of 

long-range constraints that integrate “traditional”, diamagnetic distance 

measurements in structure determination. 

[1] G. Pintacuda, N. Giraud, R. Pierattelli, A. Böckmann, I. Bertini and L. 

Emsley, Angew. Chem. Int., 2006, 46, 1079-1082. 

[2] S. Laage, R. Pierattelli, I. C. Felli, I. Bertini, A. Lesage, L. Emsley and 

G. Pintacuda, submitted. 

[3] S. Laage, J. R. Sachleben, S. Steuernagel, R. Pierattelli, G. Pintacuda, 

and L. Emsley, J. Magn. Reson., 2009, 196, 133-141. 

[4] S. Laage, A. Marchetti, J. Sein, R. Pierattelli, H. J. Sass, S. Grzesiek, 

A. Lesage, G. Pintacuda and L. Emsley, J. Am. Chem. Soc., 2008, 130, 

17216–17217. 

30 

Pa04 

Paramagnetic NMR crystallography 

Pintacuda, Guido 1 ; Kervern, Gwendal 1 ; Emsley, Lyndon 1 ; Maury, Olivier 2 

1 University of Lyon / CNRS, Centre de RMN à Très Hauts Champs, Lyon, 

France; 2 Ecole Normale Superieure de Lyon, Lyon, France 

Solid-state NMR spectroscopy can be used for structure determination 

of microcrystalline paramagnetic solids at natural isotopic abundance. 

We present a protocol which makes use of paramagnetic effects, 

measured on suitably recorded 1H NMR spectra, to define the 

conformation of a molecule in the lattice and the intermolecular packing 

in the solid phase. The method is illustrated on a family of lanthanide 

compounds. 


Relaxation 

Re01 

A unified representation of protein structural dynamics in 

solution 

Blackledge, Martin 

CEA-CNRS-UJF, Protein Dynamics and Flexibility, Institut de Biologie 

Structurale Jean-Pierre Ebel, Grenoble, France 

Proteins are inherently flexible, displaying a broad range of dynamics 

over a hierarchy of time-scales from pico-seconds to seconds. 

This molecular plasticity enables conformational changes in protein 

backbone and sidechains that are essential for biomolecular function. 

An atomic resolution description of protein flexibility is a prerequisite 

for understanding the role that structural dynamics play in biological 

processes. Although the importance of molecular flexibility is generally 

recognized, standard NMR-based structure determination protocols 

ignore the presence of protein dynamics, implying that, in common with 

X-ray crystallography, rapidly exchanging conformational equilibria are 

routinely represented in terms of a single static structure. The specific 

averaging properties of different structurally dependent parameters are 

rarely incorporated into the structure determination procedure, such 

that the resulting set of coordinates represent a poorly defined average.. 

The aim of this study is to actively use the rich dynamic information 

encoded in motionally averaged NMR parameters to develop a 

structural, dynamic and statistical mechanical molecular representation 

of the conformational behavior of proteins in solution. 

NMR residual dipolar couplings (RDCs) are uniquely sensitive to 

conformational detail and as such offer a very attractive approach to 

characterizing protein dynamics on all time scales up to the millisecond. 

The simple averaging properties of RDCs makes them amenable to 

rigorous interpretation in terms of protein structure and dynamics. 

In this work for the first time, we describe a robust procedure for 

quantitative and absolute determination of protein backbone motions 

from RDCs that requires no scaling to an external reference such as 

Lipari Szabo order parameters. We develop a novel, structure-free 

approach, based on the 3DGAF (Gaussian axial fluctuation)1,2,3 

model of peptide plane reorientation, that independently determines 

the average orientation of each peptide plane in the protein and the 

associated local conformational dynamics about this mean, in an 

entirely model free way. 4 

In a parallel study we combine accelerated molecular dynamic 

simulation5 with experimental RDCs in order to derive a molecular 

description of conformational sampling occurring up to the millisecond 

timescale, a prerequisite for interpreting all solution state experimental 

NMR data. The appropriate level of sampling is directly determined by 

comparison with dipolar and scalar couplings, again in an independent, 

but equally model-free way. The method is used to describe 

conformational dynamics occurring on timescales over many orders of 

magnitude in ubiquitin and protein G. The results reveal a remarkably 

similar amplitude and distribution of conformational dynamics compared 

to the 3DGAF analytical procedure. In particular we observe an inherent 

dependence of fast protein dynamics on local conformational topology 

sampled on slower timescales. The results provide rare insight into the 

complex hierarchy of dynamics occurring in proteins and allow us to 

develop a model of the conformational landscape native to the protein. 

These approaches therefore provide a unified structural dynamic 

representation of the statistical mechanical properties of proteins in 

solution that will provide the basis for furthering our understanding of 

molecular stability, folding and function. 

1 - Anisotropic Small Amplitude Peptide Plane Dynamics in Proteins 

from Residual Dipolar Couplings. P. Bernado and M. Blackledge. J.Am. 

Chem.Soc. 126, 4907-4920, (2004). 

2 - Identification of slow correlated motions in proteins using residual 

dipolar and hydrogen-bond scalar couplings. G. Bouvignies, P. Bernado, 

S. Meier, K. Cho, S. Grzesiek, R. Brüschweiler and M. Blackledge Proc. 

Natl. Acad. Sci. 102, 13885-13890 (2005) 

3 - Simultaneous Determination of Protein Structure and Dynamics 

using Residual Dipolar Couplings. G. Bouvignies, P.R.L.Markwick, 

R.Brüschweiler and M. Blackledge. J.Am.Chem.Soc. 128, 15100- 

15101 (2006). 

4 - Protein Conformational Flexibility from Structure-Free Analysis of 

NMR Dipolar Couplings: Quantitative and Absolute Determination of 

Backbone Motion in Ubiquitin. Loïc Salmon, Guillaume Bouvignies, 

Phineus Markwick, Nils Lakomek, Scott Showalter, Da-Wei Li, Korvin 

Walter, Christian Griesinger, Rafael Brüschweiler and Martin Blackledge 

Angewandte Chemie International Edition. 48, 4154-4157 (2009). 

5 - Exploring Multiple Timescale Motions in Protein GB3 using 

Accelerated Molecular Dynamics and NMR. P. Markwick, G. Bouvignies 

and M. Blackledge J.Am.Chem.Soc. 129, 4724-4730 (2007). 


Re02 

Investigation of correlated protein motions by the measurement 

of cross-correlated relaxation rates 

Walter, Korvin 1 ; Lee, Donghan 1 ; Lakomek, Nils-Alexander 2 ; Fenwick, 

Robert B. 3 ; Salvatella, Xavier 3 ; Becker, Stefan 1 ; Griesinger, Christian 1 

1 MPI for Biophysical Chemistry, Department of NMR based Structural 

Biology, Göttingen, Germany; 2 National Institute of Diabetes & Digestive 

& Kidney Diseases, Laboratory of Chemical Physics, Bethesda, United 

States; 3 ICREA and Institute for Research in Biomedicine, Laboratory of 

Molecular Biophysics, Barcelona, Spain 

Recently a conformational ensemble on the basis of residual dipolar 

couplings (RDCs) was presented for the protein ubiquitin (Lange & 

Lakomek et al., 2008). Former ensembles, which are based on NMR 

relaxation measurements, can represent the internal motion only up to 

the rotational correlation time, which is normally in the nanoseconds 

time range. In contrast the RDC-based ensemble can cover the time 

window up to 50 milliseconds. In addition this new ensemble covers 

all known structures, which ubiquitin adopts in complexes with other 

proteins during the recognition process. Therefore this shows that 

ubiquitin does not need the influence of the partner protein to adopt the 

required conformation (induced fit model), but has a sufficient structural 

heterogeneity for recognition of the partner proteins (conformational 

selection model). The conformational ensemble suggests a high degree 

of correlated motions. 

The experimental validation of these predictions will be addressed in the 

here presented work. Since cross-correlated relaxation (CCR) rates are 

highly sensitive to the angle between two involved dipoles, correlated 

motions should be measurable in this way. Results will be discussed in 

the presentation. 

Re03 

Global anisotropic motion in proteins in the solid state 

Lewandowski, Józef R. 1 ; Sein, Julien 1 ; Blackledge, Martin 2 ; Emsley, 

Lyndon 1 

1 Université de Lyon, CNRS / ENS-Lyon / UCB-Lyon 1, Centre de RMN 

à Très Hauts Champs, Villeurbanne, France; 2 Institut de Biologie 

Structurale Jean-Pierre Ebel CNRS/CEA/UJF, Grenoble, France 

Since the overall isotropic reorientation of macromolecules is absent 

in crystals, solid-state NMR dynamics studies of microcrystalline 

proteins have so far generally assumed that only internal motions 

make a significant contribution to nuclear spin relaxation. However, 

numerous studies employing miscellaneous techniques (and notably 

x-ray crystallography[1]) suggest that, even in the absence of overall 

ABSTRACTS ORAL


isotropic tumbling, proteins often exhibit restricted global anisotropic 

motions (AGM) – be it a small-amplitude overall reorientation or domain 

motions. In fact such collective motions often play a crucial role in 

biological activity of the proteins and are important for such processes 

as catalysis, regulatory activity, transport of metabolites etc.[2] In this 

context the ability to account for effect of AGMs on spin relaxation is 

highly desirable, as it should lead not only to more complete picture of 

protein dynamics in the solid-state but also potentially provide insight 

into a number of significant biophysical processes. 

Here we present an approach that allows to include the effect of AGMs 

on nuclear spin relaxation in solids. We use the 3-Dimensional Gaussian 

Axial Fluctuation (3D GAF)[3] model in which the motion is described 

by axial fluctuations about three orthogonal axes α, β and γ with 

amplitudes σ α , σ β and σ γ . 

We use our model to evaluate the potential impact of AGMs on the 

interpretation of 15 N longitudinal relaxation data for the microcrystalline 

protein Crh which is a dimer in this form. Our results indicate that 

presence of AGMs with amplitudes consistent with x-ray based rigidbody 

TLS[4] analysis is compatible with the experimental data and leads 

to non-negligible contributions to the relaxation rates. We suggest how 

AGMs could be included in the future analyses of the relaxation data in 

solids. 

References: 

[1] Kuriyan, J.; Weis, W. I. Proc. Nat. Acad. Sci. U.S.A. 1991, 88, 2773- 

2777. 

[2] Gerstein, M.; Lesk, A. M.; Chothia, C. Biochemistry-US 1994, 33, 

6739-6749. 

[3] Bremi, T.; Bruschweiler, R. J. Am. Chem. Soc. 1997, 119, 6672-6673. 

[4] Schomaker.V; Trueblood, Acta Crystallogr., Sect. B: Struct. Sci 1968, 

B 24, 63. 

Re04 

Water dynamics in living bacterial cells and dormant endospores 

Persson, Erik; Halle, Bertil 

Center for Molecular Protein Science, Lund University, Biophysical 

Chemistry, Lund, Sweden 

Water-biomolecule interactions have been extensively studied in dilute 

solutions, crystals, and rehydrated powders, but none of these model 

systems may capture the behavior of water in the highly organized 

intracellular milieu. Because of the experimental difficulty of selectively 

probing the structure and dynamics of water in intact cells, radically 

different views about the properties of cell water have proliferated. To 

resolve this long-standing controversy, we have measured the H spin 

relaxation rate in living bacteria cultured in D2O. Contradicting the view 

that a substantial fraction of cell water is strongly perturbed, we find 

that ≈85% of cell water in Escherichia coli and in the extreme halophile 

Haloracula marismortui has bulk-like dynamics. The remaining ≈15% of 

cell water interacts directly with biomolecular surfaces and is motionally 

retarded by a factor 15 ± 3 on average. The relaxation data also show 

that a small fraction (≈0.1 %) of cell water exchanges from buried 

hydration sites on the microsecond time scale. 

Whereas 70% of the cell volume in E. coli is occupied by water, this 

number is only 30% in the core of a dormant bacterial spore. In such 

dry milieu, the water dynamics must play an extremely important role. 

We were able to selectively probe for the water dynamics within and 

outside the core of intact spores. The water molecules present in the 

core have similar motional perturbation as in the hydration layer of 

biomolecules in the cytoplasm of E. coli. The relaxation data also reveal 

that the water permeation across the core inner membrane is orders of 

magnitude lower than for a membrane made from E. coli lipid extract 

but this low water permeability is not critical for the spore wet heat 

resistance. The low permeability may on the other hand be critical for 

the spore’s resistance against toxic chemicals. 

32 

Small Molecules 

Sm01 

Conformational changes and biological activities: From small 

molecules to large biomolecular complexes 

Kessler, Horst 

TU München, Insitute for Advanced Study Dept Chemie, Garching, 

Germany 

The procedure of conformational analysis by NMR and MD is presented 

and possible pitfalls are discussed. Recently it turned out that more and 

more unreliable structure determinations of peptides are reported due 

to uncritical application of routine protein NMR technology to peptides, 

neglecting the distinct differences between small and large molecules. 

Conformational transitions are also important for proteins. Naturally 

unfolded protein domains in p53 and in constant domains of antibodies 

are important features in understanding the processing and regulation 

of protein functions. These structural studies help to explain differences 

in fibril formation, involved in several folding diseases. Finally, structure 

determination of the C-terminal domain of spider silk gives insight into 

the regulation of the spinning process. 

All these different processes have in common their conformational 

transitions and their regulation via surrounding effects (solvent, 

temperature, salt concentration, pH, interaction with biological partner 

molecules such as DNA, other proteins and chaperons). 

Sm02 

Configurational assignment of complex natural products by 

NMR and distance geometry; application to pyrrole-imidazole 

alkaloids 

Köck, Matthias; Schmidt, Gesine 

Alfred-Wegener-Inst. for Marine and Polar Research, Bremerhaven, 

Germany 

The determination of the relative and absolute configuration of natural 

products is essential to understand their biological activity. Methods 

as X-ray crys¬tallography requires crystalline products and chemical 

synthesis is usually very time consuming and by no means reliable. 

Here we want to discuss how effective NOE/ROE effects in combination 

with computational methods can be used for this purpose. The NOE/ 

ROE restraints may be used in a qualitative way or as restraints in EM 

or MD simulations. These approaches are problematic with a large 

number of unknown stereogenic centers. Therefore, a method is 

required which allows the determination of all unknown stereogenic 

centers simultaneously and without the necessity of crystalline 

products. Distance geometry (DG) in combination with distance bounds 

driven dynamics (DDD) calculations using interproton distances and 

floating chirality are discussed here for the determination of the relative 

configuration (floating chirality restrained DG/DDD, fc-rDG/DDD). 

The fc-rDG/DDD method is applied to structurally very complex 

members of the pyrrole-imidazole alkaloid family. The investigated 

molecules have all 8 stereogenic centers in common. The results of 

these calculations on tetrabromostyloguanidine (1) revised the relative 

configuration of the palau’amine congeners in 2007. Here we expand 

these investigations on axinellamine A (2) and new derivatives of 

massadine chloride (3). 

1 a) R. Kaptein, R. Boelens, R. M. Scheek, W. F. van Gunsteren, 

Biochemistry 1988, 27, 5389–5395; b) R. M. Scheek, W. F. van 

Gunsteren, R. Kaptein, Methods Enzymol. 1989, 177, 204–218. 

2 a) P. L. Weber, R. Morrison, D. Hare, J. Mol. Biol. 1988, 204, 483– 

487; b) T. A. Holak, D. Gondol, J. Otlewski, T. Wilusz, J. Mol. Biol. 1989, 


210, 635–648. 

3 a) M. Reggelin, M. Köck, K. Conde-Frieboes, D. F. Mierke, Angew. 

Chem. Int. Ed. 1994, 33, 753–755; b) M. Köck, J. Junker, J. Mol. 

Model. 1997, 3, 403–407; c) M. Köck, J. Junker, J. Org. Chem. 1997, 

62, 8614–8615. 

4 a) A. Grube, M. Köck, Angew. Chem. Int. Ed. 2007, 46, 2320–2324; 

b) M. S. Buchanan, A. R. Carroll, R. Addepalli, V. M. Avery, J. N. A. 

Hooper, R. J. Quinn, J. Org. Chem. 2007, 72, 2309–2317; c) H. 

Kobayashi, K. Kitamura, K. Nagai, Y. Nakao, N. Fusetani, R. W. M. van 

Soest, S. Matsunaga, Tetrahedron Lett. 2007, 48, 2127–2129. 

5 S. Urban, P. de A. Leone, A. R. Carroll, G. A. Fechner, J. Smith, J. N. A. 

Hooper, R. J. Quinn, J. Org. Chem. 1999, 64, 731–735. 

6 A. Grube, S. Immel, P. S. Baran, M. Köck, Angew. Chem. Int. Ed. 2007, 

46, 6721–6724. 

Sm03 

Conformational study on the tripeptide 

Hyp(OBn)–β-ACC–Pro–OBn in solution by NMR and MD 

Fleischmann, Matthias 1 ; Schmid, Markus 1 ; D‘Elia, Valerio 1 ; Reiser, 

Oliver 1 ; Gronwald, Wolfram 2 ; Gschwind, Ruth Maria 1 

1 University of Regensburg, Institute of Organic Chemistry, Regensburg, 

Germany; 2 University of Regensburg, Institute of Functional Genomics, 

Regensburg, Germany 

In biological systems and organocatalysis specific conformations of 

peptides often provide high activities and selectivities. In this context, 

derivatives of the unnatural cis-β-aminocyclopropanecarboxylic acid 

(β-ACC) have been shown to stabilize secondary structure elements 

even in short peptide sequences.[1] Therefore, foldamers containing 

β-ACCs are used in medicinal chemistry as analogues for the 

neuropeptide Y with high selectivity for the Y1 receptor.[2] Additionally, 

tripeptides with β-ACC and proline like residues showed organocatalytic 

activity in inter- and intramolecular aldol reactions yielding high 

diastereo- and enantioselectivities.[3] In a previous conformational 

study on Pro–Pro–(-)-β-ACC–OBn 1 and Pro–(-)-β-ACC–Pro–OBn 2 

both tripeptides showed limited flexibility in solution,[4] which is crucial 

for the 

investigation of mechanistic details and active conformations of 

organocatalysts in general. 

Here, we present the structure investigation of Hyp(OBn)–(+)-β- 

ACC–Pro–OBn 3, which is a further development of 1 and 2. To stabilize 

intramolecular interactions in this ultrashort peptide, the NMR studies 

of 3 were performed in CDCl 3 at 240 K. The NOESY distance restraints 

were refined by a spindiffusion relaxation matrix calculated with 

AUREMOL and molecular dynamics simulations were done with CNS. 

[1] S. De Pol, C. Zorn, C.D. Klein, O. Zerbe, O. Reiser, Angew. Chem. Int. 

Ed. 

2004, 43, 511-514. 

[2] N. Koglin, C. Zorn, R. Beumer, C. Cabrele, C. Bubert, N. Sewald, O. 

Reiser, 

A. Beck-Sickinger, Angew. Chem. Int. Ed. 2003, 42, 202-205. 

[3] V. DÉlia, H. Zwicknagl, O. Reiser, J. Org. Chem. 2008, 73, 3262. 

[4] M. Schmid, M. Fleischmann, V. DÉlia, O. Reiser, W. Gronwald, R. M. 

Gschwind, 

ChemBioChem. 2009, 10, 440-444. 


Sm04 

Determination of structure and dynamics of an organocatalyst 

using residual dipolar coupling 

Thiele, Christina; Heiles, Sven 

Technische Universität Darmstadt, Clemens Schöpf Institut, Darmstadt, 

Germany 

Residual Dipolar Couplings (RDCs) are becoming increasingly important 

not only in biomacromolecular NMR but also in the determination 

of configuration and conformation of small organic compounds [1]. 

Methods are being developed to also include conformational flexibility 

into these analyses [2]. So far, however, RDCs have not been applied to 

gain insight into reaction mechanisms. 

There is a huge discrepancy between the enormous importance of 

catalytic processes in organic synthesis and the knowledge on the 

structure and dynamics of the catalytically active species. This is also 

true for the photoswitchable organocatalyst investigated here. 

The form, which was expected to be catalytically inactive (shielded 

lone pair on nitrogen) exhibited some unexpected pseudo-background 

reactivity. The reason for this background reactivity could be clarified 

from an analysis of the conformation and dynamics obtained from 

residual dipolar couplings measured in high molecular weight PBLG[3]. 

References: 

[1] Reviews: C. M. Thiele, Eur. J. Org. Chem., 2008, 5673-5685; C. M. 

Thiele, Conc. Magn. Res. 2007, 30A, 65-80. 

[2] C. M. Thiele, A. Marx, R. Berger, J. Fischer, M. Biel, A. Giannis, 

Angew. Chem. Int. Ed. 2006, 45, 4455-4460; A. Schütz, J. Junker, 

A. Leonov, O. F. Lange, T. F. Molinski, C. Griesinger, J. Am. Chem. Soc. 

2007, 129, 15114-15115. 

[3] A. Marx, C.M. Thiele, Chem. Eur. J. 2009, 15, 254–260. 

[4] R. S. Stoll, M. V. Peters, A. Kühn, S. Heiles, R. Goddard, M. Bühl, C. 

M. Thiele, S. Hecht, J. Am. Chem. Soc. 2009, 131, 357-367. 

ABSTRACTS ORAL


34 

Solid State NMR 

Sn01 

Structural insights into Ure2p full-length prion fibrils by solidstate 

NMR spectroscopy 

Loquet, Antoine 1 ; Bousset, Luc 2 ; Gardiennet, Carole 1 ; Sourigues, 

Yannick 2 ; Wasmer, Christian 3 ; Habenstein, Birgit 1 ; Schuetz, Anne 3 ; Meier, 

Beat H. 3 ; Melki, Ronald 2 ; Böckmann, Anja 1 

1 IBCP UMR 5086 CNRS/Université de Lyon 1, Lyon, France; 2 LEBS UPR 

3082 CNRS, Gif/Yvette, France; 3 ETH Zurich, Physical Chemistry, Zurich, 

Switzerland 

Protein misfolding and subsequent aggregation is the hallmark of over 

30 diseases termed “conformational” diseases; the heterogeneity of 

the high molecular weight protein oligomers and their molecular mass 

make them particularly not suitable for structural studies at atomic 

resolution by classical methods as x-ray crystallography and solution 

NMR. Moreover, it has long been believed that misfolded proteins like 

prions and amyloids give poorly resolved NMR spectra. 

We show that the spectra of full-length fibrils of the 40.5 kDa Ure2 

yeast prion protein generated under physiologically relevant conditions 

interestingly lead to very highly resolved solid-state NMR spectra. 

Prion fibrils formed under physiological conditions are therefore highly 

ordered objects on the molecular level. Ure2p is a two-domain protein 

made of a flexible N-terminal prion domain that spans residues 1-93 

and of a compactly folded C-terminal domain that spans residues 94- 

354, with known crystal structure.[1] The N-terminal domain is critical 

for the assembly of Ure2p into protein fibrils and assembles when 

isolated into typical amyloids. Comparing the full-length NMR spectra 

with the corresponding spectra of the prion and globular domains in 

isolation reveals that the globular part, in particular, shows almost 

perfect structural order. For the prion domain, the spectra reflect partial 

disorder suggesting structural heterogeneity, both in isolation and in fulllength 

Ure2p fibrils, although to different extents. The spectral quality 

is surprising in the light of existing structural models for Ure2p, and in 

comparison to the corresponding spectra of the only other full-length 

prion fibrils (HET-s) investigated so far. Our findings provide insight into 

the structural diversity underlying prion propagation suggesting that no 

unique mode exists for the assembly of these proteins into fibrils. 

[1] L. Bousset, H. Belrhali, J. Janin, R. Melki, S. Morera, Structure 2001, 

9, 39. 

Sn02 

Determination of a molecular torsional angle in the 

bathorhodopsin photointermediate of rhodopsin by doublequantum 

solid-state NMR 

Concistre, Maria 1 ; McLean, Neville 1 ; Johannessen, Ole G. 1 ; Pileio, 

Giuseppe 1 ; Bovee-Geurts, Petra H. M. 2 ; Brown, Richard C. D. 1 ; De Grip, 

Willem J. 2 ; Levitt, Malcolm H. 1 

1 University of Southampton, School of Chemistry, Southampton, United 

Kingdom; 2 Radboud University, Nijmegen Centre for Molecular Life 

Sciences, Nijmegen, Netherlands 

Rhodopsin is a G-protein coupled receptor (GPCR) responsible for dim 

light vision in mammals. Light absorption isomerizes its retinylidene 

chromophore to form the strained all-E intermediate bathorhodopsin, 

which has been studied by X-ray diffraction [1], femtosecond stimulated 

Raman spectroscopy [2], and solid-state NMR [3]. 

We recently studied bathorhodopsin by low-temperature solid-state 

NMR, using in situ illumination of the sample in a customized NMR 

probe. A solid state NMR method called double-quantum heteronuclear 

local field spectroscopy (2Q-HLF) has been developed for the direct 

estimation of torsional angle in a 1H-13C-13C-1H molecular fragment 

[4]. 

We applied 2Q-HLF spectroscopy to the first intermediate of the 

rhodopsin photosequence in order to characterize the changes in 

structure of the chromophore during the initial stages of the signal 

transduction after photoactivation. The results indicate a deviation from 

the planarity in bathorhodopsin of at least 40° about the C11=C12 

double bond. This conclusion is in agreement with previous Raman 

observations [2]. 

References: 

[1] Nakamichi, H.; Okada, T. Angew. Chem. Int. Ed. 2006, 45, 4270- 

4273. 

[2] Kukura P.; McCamant, D. W.; Yoon, S.; Wandschneider, D. B; 

Mathies, R. A. Science 2005, 310, 1006-1009. 

[3] Concistrè, M.; Gansmüller, A.; McLean, N.; Johannessen, O. G.; 

Marín Montesinos, I.; Bovee-Geurts, P. H. M.; Verdegem, P.; Lugtenburg, 

J.; Brown, R. C. D.; DeGrip, W. J.; Levitt, M. H. J. Am. Chem. Soc. 2008, 

130, 10490-10491. 

[4] Feng, X.; Lee, Y. K.; Sandström, D.; Edén M.; Maisel, H.; Sebald, A.; 

Levitt, M. H. Chem. Phys. Lett. 1996, 257, 314-320. 

Sn03 

NMR with small magnets and well-defined fields 

Blümich, Bernhard; Casanova, Federico; Danieli, Ernesto; Perlo, Juan 

RWTH-Aachen, ITMC/MC, Aachen, Germany 

NMR with small magnets can be conducted outside the NMR lab to 

which high-field magnets are banned [1]. Two types of such magnets 

are available, Halbach-type magnets which accommodate the sample 

inside and stray-field magnets which accommodate the sample outside 

[2]. Early magnets of either kind were of simple design with little control 

over a customized definition of the magnetic-field profile [1]. With the 

advent of the Profile NMR-MOUSE [3] and the spectroscopy NMR- 

MOUSE [4] new shim technologies were developed to adjust the field 

generated by permanent magnets of varying perfection in shape and 

granular structure [5], for example, to uniformity in the gradient or in the 

field. Such magnets can be built small and compact with a favorable 

ratio of sensitive volume to magnet size. Similar to well-logging 

instruments they can be moved to the site of interest and operated 

under extreme conditions of pressure and temperature. The state of the 

art will be addressed [6-8] and novel magnets and their uses will be 

presented. 

Sn04 

Optimal control in NMR spectroscopy: Design your own 

experiment 

Tosner, Z. 1 ; Kehlet, C. 2 ; Nielsen, A.B. 2 ; Bjerring, M. 2 ; Nielsen, J.T. 2 ; 

Straasoe, L.A. 2 ; Vinding, M.S. 2 ; Vosegaard, T. 2 ; Khaneja, N. 3 ; Glaser, 

S.J. 4 ; Nielsen, N.C. 2 

1 Charles University in Prague, Dept. of Chemistry, Faculty of Science, 

Prague, Czech Republic; 2 University of Aarhus, Dept. of chemistry, 

inSPIN, iNANO, Aarhus, Denmark; 3 Harvard University, Division of 

Applied Sciences, Cambridge, United States; 4 Technische Universitat 

Munchen, Dept. of Chemistry, Munchen, Germany 

More than a dozen of papers has already demonstrated the use of 

optimal control theory (OCT) for experiment design within the field of 

liquid-state NMR, solid-state NMR and magnetic resonance imaging. 

OCT offers a powerful optimization strategy that can efficiently handle 

hundreds-to-thousands of variables to exploit the full degree of 

experimental freedom and thereby can provide optimal experiments that 


may be difficult to reach using standard analytical design methods. 

In our presentation we demonstrate the implementation of optimal 

control into the open source simulation package SIMPSON that has 

been recently released. The software allows efficient development and 

optimization of NMR experiments for a wide range of applications, 

including liquid and solid state NMR, magnetic resonance imaging, 

quantum computation and combinations between NMR and other 

spectroscopies. It enables optimization of experiments with considering 

rf and static field inhomogeneities, variations of spin system parameters 

or limitations of experimental resources, as well as reduction of rf field 

strengths (e.g. to avoid sample heating). We provide a brief account 

on relevant theory and describe in particular the simple computational 

interface to use optimal control routines in SIMPSON. We wish to 

show that using SIMPSON everyone can now design their own pulse 

sequences that meet the specific needs of a particular laboratory. 

The second part of the presentation introduces various new experiments 

designed using optimal control theory with focus on dipolar recoupling 

in biological solid state NMR, broadband pulses for liquid and solid state 

NMR, and volume selective excitation in magnetic resonance imaging. 

Reference: 

Z. Tošner, T. Vosegaard, C.T. Kehlet, N. Khaneja, S.J. Glaser, N.C. 

Nielsen, J. Magn. Reson., 197, 2009, 120-134 

Solution NMR 


So01 

Limits on control of spin dynamics 

Khaneja, Navin 

Harvard University, Cambridge, United Kingdom 

Some questions of fundamental interest to a NMR spectroscopist 

include, what is the shortest pulse sequence that produces a desired 

unitary transformation in a coupled spin system. How much coherence 

or polarization can be transferred between coupled spins in the 

presence of relaxation? What are the optimal pulse sequences that 

achieve these limits. We detail some recent developments that provide 

concrete answers to many of these problems, and describe applications 

to solution and solid state NMR. We present some new techniques 

for design of NMR experiments in the presence of Larmor dispersion, 

and inhomogeneous static and rf-fields. Finally, we present some new 

heteronuclear and homonuclear decoupling methods. 

So02 

Prochiral and chiral discrimination in NMR spectra: solutes in 

variably stretched and compressed anisotropic gels 

Naumann, Christoph; Kuchel, Philip W. 

University of Sydney, School of Molecular and Microbial Biosciences, 

Sydney, Australia 

We demonstrate prochiral and chiral spectral resolution using residual 

2H NMR quadrupolar splittings and residual dipolar couplings in 1H 

and 13C NMR over a wide range of anisotropic conditions in liquid 

samples. We use a reversible gel-stretching/compressing device in 

a conventional high field NMR spectrometer. We show the stability 

of the gels as well as their unique ability to switch between multiple 

sequentially stretched and compressed states, thus changing the sign 

of residual dipolar couplings in 1H and 13C NMR spectra. This flexibility 

is important for resolving spectra of mixtures of chiral compounds and 

for structure determination of selected peptides. 

The method is based on gels carrying polar guest molecules in an 

elastic silicone rubber tube; this enables the rapid and reversible 

adjustment of the extent of nuclear alignment inside NMR samples by 

merely stretching or relaxing the silicone rubber tube.1-4 Each sample 

is easily and quickly adjusted several times, leading to spectra with the 

guest molecules exposed to different extents of anisotropy. This is an 

important advance on previous work in liquid crystals5, and stressed 

polyacrylamide gels6, because generally not all extents of alignment 

resolve all mixtures. The chirality of these gels means that not only 

enantiomers but also prochiral sites may be differentiated once the gel 

is stretched or compressed. 

References: 

1. Naumann, C. and Kuchel, P. W. (2008) J. Phys. Chem. A, 112, 8659- 

8664. 

2. Naumann, C.; Bubb, W. A.; Chapman, B. E.; Kuchel, P. W. (2007) J. 

Am. Chem. Soc., 129, 5340-5341. 

3. Kuchel, P. W.; Chapman, B. E.; Mueller, N.; Bubb, W. A.; Philp, D. J.; 

Torres, A. M. (2006) J. Magn. Reson., 180, 256-265. 

4. Kuchel, P. W.; Naumann, C. (2008) J. Magn. Reson., 192, 48-59. 

5. For example a) Kay, L. E.; Thomson, D. S.; Prestegard, J. H. (1988) 

Magn. Reson. Chem., 26, 860-866; b) Weise, C. F.; Weisshaar, J. C. 

(2003) J. Phys. Chem. B, 107, 3265-3277; c) Canet, I.; Courtieu, J.; 

Loewenstein, A.; Meddour, A. ; Pechine, J. M. (1995) J. Am. Chem. Soc., 

117, 6520-6526. 

6. Jones, D. H.; Opella, S. J. (2004) J. Magn. Reson., 171, 258-269. 

ABSTRACTS ORAL


So03 

High resolution NMR in inhomogeneous or fluctuating magnetic 

fields 

Pelupessy, Philippe 1 ; Rennella, Enrico 2 ; Bodenhausen, Geoffrey 1 

1 ENS, Department of Chemistry, Paris, France; 2 Università degli Studi di 

Udine, Department of Biomedical Sciences & Technologies, Udine, Italy 

Nuclear magnetic resonance experiments are usually carried out 

in homogeneous magnetic fields. In many cases however, it is not 

practical to insert the sample into the bore of a magnet, so that ex 

situ measurements are the only option. In other cases, the inherent 

heterogeneities of the materials or (possibly living) samples under 

investigation make it virtually impossible to achieve homogeneous 

fields. Finally, resistive or hybrid magnets can generate very high fields 

(currently up to 45 T or ~2 GHz for proton NMR), but these fields 

fluctuate because of unstable power supplies and vibrations arising 

from cooling. Here we show how high resolution NMR spectra can be 

obtained in very inhomogeneous static fields with unknown field profiles 

and unknown slow fluctuations. The novel approach combines ‘ultrafast’ 

single-scan methods with zero-quantum spectroscopy. 

So04 

Towards real-time NMR spectroscopy of high molecular weight 

proteins 

Amero, Carlos 1 ; Schanda, Paul 2 ; Gans, Pierre 1 ; Brutscher, Bernhard 1 ; 

Boisbouvier, Jerome 1 

1 Institut de Biologie Structurale Jean-Pierre Ebel CNRS-CEA-UJF, 

Grenoble, France; 2 Laboratorium für Physikalische Chemie ETH, Zurich, 


Real-time NMR spectroscopy have been applied successfully to small 

proteins over the past decade. We recently introduced an optimized 

NMR experiment that make similar real-time studies feasible for large 

molecular weight proteins. The SOFAST-methyl-TROSY experiment 

combines the advantages of methyl-TROSY and SOFAST-HMQC, 

to record high quality methyl 1H-13C correlation spectra of protein 

assemblies of several hundreds of kDa in few seconds. This technique 

may prove useful for real-time investigation of macromolecular 

folding and self-assembly. Here we present applications of this new 

method for the TET2 protein, an aminopeptidase involved in peptide 

degradation in the Archaea bacterium Pyrococcus horikoshii, forming a 

homododecamer of 468 kDa. 

36 

Solid State Pysics 

Sp01 

The use of quadrupole NMR spectroscopy for the investigation 

of intermetallic compounds 

Haarmann, Frank 

RWTH Aachen, Institute of Inorganic Chemistry, Aachen, Germany 

Intermetallic compounds are of huge technological interest since 

they feature physical and chemical properties like magnetism or 

superconductivity or resistance against corrosion even at high 

temperatures. A straight forward synthesis of these materials with the 

required properties is currently not possible due to the leakage of the 

understanding of the chemical bonding of intermetallic compounds. 

A second frequently observed feature of intermetallic compounds is 

disorder. Therefore, tools are needed to get insights into the chemical 

bonding and the local arrangements of the atoms. NMR spectroscopy 

seems to be highly suited for this purpose. 

Systematic studies on model systems MGa 2 with M = Ca, Sr, Ba 

and MGa 4 with M = Na, Ca, Sr, Ba show that by a combination of 

NMR spectroscopy and quantum mechanical calculations valuable 

information about chemical bonding can be obtained. Perfect 

agreement of the EFG determined from NMR experiments and quantum 

mechanical calculations was obtained. An analysis of the contributions 

of the EFG indicates a large difference of the charge distribution in 

the vicinity of the Ga atoms of structurally related compounds. This is 

caused by a variation of the population of the p x ,p y ,p z -like states of the 

Ga atoms. In addition, it could be shown by these investigations that 

the structure models for SrGa 2 and BaGa 2 have to be modified. Both 

compounds were known from X-ray diffraction to crystallize in the 

AlB 2 -type of structure with planar honeycomb like nets of Ga atoms [1]. 

NMR spectroscopy and quantum mechanical investigations evidence a 

puckering of the Ga layers by only +/- 0.125 [2]. 

Additional examples like disorder in Cu 1-x Al 2 [3] and other documenting 

the advantage of quadrupole NMR spectroscopy on intermetallic 

compounds will be presented. 

[1] G. Bruzzone; Boll. Sci. Fac. Chim. Ind. Bologn. (1966) 24, 113. 

[2] F. Haarmann, K. Koch, D. Grüner, W. Schnelle, O. Pecher, R. Cardoso- 

Gil, H. Borrmann, H. Rosner, Yu. Grin; Chem. Eur. J. (2009) 15(7) 1673. 

[3] F. Haarmann, M. Armbrüster, Yu. Grin; Chem. Mater. (2007) 19, 

1147. 

Sp02 

Magnetic resonance study of detonation nanodiamonds with 

surface modified by transition metal ions 

Panich, Alexander 1 ; Shames, Alexander 1 ; Osipov, Vladimir 2 ; Alexenskii, 

Alexander 2 ; Vul’, Alexander 2 

1 Ben-Gurion University of the Negev, Department of Physics, Be’er- 

Sheva, Israel; 2 Ioffe Physico-Technical Institute, St. Petersburg, Russian 

Federation 

We report on NMR and EMR study of detonation nanodiamonds 

(DND) with surface chemically modified by copper and cobalt ions. 

Measurements of the 13C nuclear spin-lattice relaxation rate R1 of 

the diamond core carbons show stretched exponential magnetization 

recovery and 3 orders of magnitude increase in R1 in comparison with 

the natural diamond, attributed to the interaction of nuclear spins with 

unpaired electron spins of dangling bonds. Modification of the DND 

surface by Cu and Co causes an additional increase in R1, indicating 

appearance of paramagnetic Cu2+ and Co2+ complexes at the DND 

surface and their interaction with the carbon nuclear spins, both directly 


and via a coupling of Cu2+ and Co2+ electron spins with electron 

spins of the carbon-inherited defects. Subsequent annealing of the 

modified samples in the hydrogen flow at 550 and 900 oC results in 

an inverse process, i.e., a reduction of the relaxation rate, indicating 

that upon annealing these complexes are destroyed, and metal ions 

presumably join each other forming copper and cobalt nanoclusters. 

Co nanoclusters are ferromagnetic, which results in the noticeable 

broadening of 13C NMR line. 

EMR spectrum of pure DND sample shows intensive singlet originating 

from carbon-inherited defects, while the spectra of cooper- and 

cobalt- modified samples reveal pronounced hyperfine structure due 

to a coupling of electron spin with the spins of 63,65Cu (I = 3/2) and 

59Co (I = 7/2) nuclei, indicating the appearance of magnetically diluted 

mononuclear Cu2+ and Co2+ ion complexes. Increase in the Cu or 

Co concentration yields progressive increase of the corresponding 

additional EMR signals as well as to the consequent broadening of 

the intensive carbon-inherited EMR line. Annealing of the coppermodified 

samples at 550 and 900 oC causes an inverse process - line 

narrowing. Thus our NMR and EMR data correlate with each other and 

allow shading light on interactions and possible locations of transition 

metal ions on the nanodiamond surface. 

Sp03 

Low-temperature NMR and quantum theory of dihydrogen 

endofullerenes 

Mamone, Salvatore 1 ; Ge, Min 2 ; Huvonen, D. 2 ; Nagel, U. 2 ; Danquigny, 

Alain 1 ; Cuda, Francesco 1 ; Grossel, Martin 1 ; Murata, Yasujiro 3 ; Komatsu, 

Koichi 3 ; Levitt, Malcolm 1 ; Rõõm, Toomas 2 ; Carravetta, Marina 1 

1 Southampton University, School of Chemistry, Southampton, United 

Kingdom; 2 National Institute of Chemical Physics and Biophysics, Tallinn, 

Estonia; 3 Institute for Chemical Research, Kyoto, Japan 

The dihydrogen endofullerene H2@C60 is an interesting example 

of a trapped quantum rotor which has been approached both by 

experimental and theoretical methods. We have obtained lowtemperature 

NMR data down to 1.7 Kelvin, including proton lineshapes 

and relaxation measurements, on a set of dihydrogen endofullerenes 

with a variety of cage symmetries. This data has been combined 

with infrared spectroscopy and inelastic neutron scattering to obtain 

a detailed picture of the quantum dynamics of the confined rotor. In 

this talk an analytical treatment of H2@C60 quantum dynamics from 

first quantum principles is presented. The energy level structure of 

the vibrating rotor is expressed in terms of a few parameters which 

describe the deviations of the effective potential from a spherical 

harmonic confinement. The theory is intended to allow the NMR, IR 

and INS data to be understood in a unified formalism. Successes of the 

theory will be presented, as well as those cases for which we still have 

an incomplete understanding of the NMR behaviour. 


Sp04 

Antiferromagnetic insulator parent state of Cs 3 C 60 

superconductor 

Jeglic, Peter 1 ; Ganin, Alexey Y. 2 ; Takabayashi, Yasuhiro 3 ; Rosseinsky, 

Matthew J. 2 ; Prassides, Kosmas 3 ; Arcon, Denis 4 

1 Institute “Jozef Stefan”, Ljubljana, Slovenia; 2 University of Liverpool, 

Liverpool, United Kingdom; 3 University of Durham, Durham, United 

Kingdom; 4 Institute “Jozef Stefan”, University of Ljubljana, Ljubljana, 

Slovenia 

The well studied face-centered cubic A 3 C 60 (A = Na, K, Rb, Cs) are all 

superconducting with superconducting transition temperatures (T C ’s) up 

to 33 K. Their T C ’s increase monotonically with interfulleride separation 

in accordance with the Bardeen-Cooper-Schrieffer (BCS) theory. 

Recently, interest in A 3 C 60 has been revived after the discovery of 

superconductivity in the body-centered cubic A15 Cs 3 C 60 phase at T C 

= 38 K [1]. Non-monotonic dependence of T C with pressure, where T C 

first increases and then decreases with increasing pressure (decreasing 

interfulleride separation) cannot be explained within BCS theory. We 

have shown by 133 Cs and 13 C NMR [2] that the superconductivity state 

emerges directly from the antiferromagnetic insulating state with the 

application of pressure. This highlights the importance of electronic 

correlations in A15 Cs 3 C 60 , which are crucial for complex phase 

diagrams in many electronic systems, including high-temperature 

superconductors. 

References 

[1] A. Y. Ganin et al., Nat. Mater. 7, 367 (2008). 

[2] Y. Takabayashi, A. Y. Ganin, P. Jeglic, D. Arcon, T. Takano, Y. Iwasa, Y. 

Ohishi, M. Takata, N. Takeshita, K. Prassides, M. J. Rosseinsky, Science 

323, 1585 (2009). 

ABSTRACTS ORAL


38 

Transport and Diffusion 

Td01 

Association of charged entities as observed by diffusion and 

electrokinetic NMR 

Furó, István 

Royal Institute of Technology, Department of Chemistry, Stockholm, 

Sweden 

When ions, molecules, and particles associate, their characteristic 

translational mobility and their apparent charge change. Both of these 

properties can be accurately measured by two members of the family 

(or, rather, tribe) of NMR methods. One of those - diffusion NMR - has 

been with us for many years; some novel applications of it to ion pairing 

in ionic thermotropic liquid crystals and to aggregation of charged clay 

particles will be presented. The other technique - electrophoretic NMR 

or, perhaps in a broader sense, electrokinetic NMR (eNMR) - is not 

necessarily much younger but has remained less developed during the 

past decades. Methodological improvements for eNMR experiments are 

presented with emphasis on improved signal-to-noise, accuracy, and 

accessibility and ease-of-use. The improved methodology is illustrated 

in applications to ion pairing and inclusion complexes. In the more 

applied direction, electrokinetic transport in fuel cell membranes will be 

assessed. 

Td02 

Fast q-space acquisition to monitor red blood cell shape 

evolution 

Pages, Guilhem 1 ; Larkin, Timothy 1 ; Szekely, David 2 ; Torres, Allan 3 ; 

Kuchel, Philip 1 

1 The University of Sydney, School of Molecular and Microbial 

Biosciences, Sydney, Australia; 2 Victor Chang Cardiac Research 

Institute, Division of Molecular Cardiology and Biophysics, Sydney, 

Australia; 3 The University of Western Sydney, Nanoscale Organisation 

and Dynamics Group, Sydney, Australia 

Restricted diffusion of water inside red blood cells (RBCs) can be 

monitored by pulsed field-gradient stimulated-echo (PFGSTE) NMR 

spectroscopy. The data yield a plot of the signal vs the spatial wavenumber 

vector q that characterizes the mean cell shape. Normal human 

RBCs have a characteristic biconcave disc shape and they align with 

B 0 , hence the diffusion-diffraction pattern exhibits minima and maxima 

at particular q values. Application of a digital filter [1] to the NMR 

data yields a mean-displacement spectrum. PFGSTE NMR enables 

experiments under conditions close to in vivo (high hematocrit) but has 

suffered from long experiment times (~ 1 h). 

To decrease the acquisition time of q-space plots and thus investigate 

RBC shape evolution, we implemented a bipolar stimulated-echo pulse 

sequence based on unbalanced gradient pulse pairs [2]; this requires 

only two transients per gradient magnitude. A difference between the 

minima in the q-space plots, between both classical and fast-acquisition 

PFGSTE experiments was observed. We demonstrated experimentally 

and theoretically that the short gradient pulse approximation is not 

applicable under our experimental conditions. 

RBCs were treated with NaF to induce the evolution from discocytes 

to spherocytes via various types of echinocytes (spherical shapes with 

rounded and then spiky protrusions). We compared PFGSTE data with 

light-microscope pictures and 31 P NMR estimated ATP concentration [3]. 

PFGSTE experiments were also alternatively run between two gradient 

axes: along z and y. The overlapping of NMR signal decays during the 

time course demonstrated the formation of spherical RBCs. In addition, 

we depleted cells of Mg 2+ and quantified the kinetics of reversion, from 

echinocytes to discocytes, after the reintroduction of this ion. 

In conclusion, our approach allows the study of the energetics and 

kinetics of RBC shape changes under conditions that are close to 

physiological with a temporal resolution that is necessary to assess the 

concurrent metabolic state of the cells. 

1. Kuchel P.W. et al. Magn Reson Med 52 907 

2. Pelta M.D. et al. Magn Reson Chem 40 S147 

3. Pages G. et al. J Magn Reson Imag 28 140 

Td03 

Water exchange over the cell membrane 

Åslund, Ingrid; Nowacka, Agnieszka; Nilsson, Markus; Topgaard, Daniel 

Lund University, Physical Chemistry, Lund, Sweden 

Water transport over the cell membrane is a crucial process for the 

function of living cells. The diffusional membrane permeability refers 

to a stationary system in which there is no net flux of water. Apparent 

diffusion coefficients (ADCs) as measured by pulsed gradient spin 

echo (PGSE) NMR can be used to resolve the NMR signal of intra- and 

extracellular water. The Diffusion EXchange SpectroscopY (DEXSY) 

technique [1] comprises two PGSE blocks separated by a variable 

mixing time. The gradients of each PGSE block are incremented 

independently and the resulting 2D data set is subjected to a 2D 

inverse Laplace transform (ILT) yielding a 2D diffusion spectrum. The 

appearance of cross peaks is a signature of molecular exchange 

between component with different ADC such as intra- and extracellular 

water. Although conceptually appealing, the practical usefulness of 

DEXSY is limited by the long experiment time required to probe two 

diffusion and one mixing time dimension. 

In this work we suggest performing the DEXSY experiment in a way 

analogous to the Goldman-Shen experiment [2] where a filter block 

and a mixing time precede signal detection. Different components 

are resolved based on differences in linewidth and the filter block 

is optimized for removing the broad components of the spectrum. 

With increasing mixing time the amplitude of the various components 

approach their equilibrium values. The Filter EXchange SpectroscopY 

(FEXSY) technique is obtained by replacing the first variable gradient 

PGSE block of the DEXSY experiment with a fixed gradient filter block 

optimized to remove the signal from molecules having a large ADC. 

This «diffusion filter» corresponds to the dipolar filter of the Goldman- 

Shen experiment. In comparison to DEXSY the dimensionality of the 

experiment is reduced, resulting in drastically shorter experiment times. 

FEXSY is demonstrated by measuring the cell membrane permeability of 

baker’s yeast at a time resolution of a few minutes. Yeast is a common 

model system in cell biology for studies of structure and biochemical 

process in eukaryotic cells. 

[1] Callaghan, Furo. J. Chem. Phys. (2004). 

[2] Goldman, Shen. Phys. Rev. (1966). 


Td04 

Can a self-diffusion spectrum of water, as measured by a novel 

NMR technique, reveal any wet secrets? 

Stepišnik, Janez 1 ; Serša, Igor 2 ; Mohoriè, Aleš 1 

1 University of Ljubljana, FMF, Ljubljana, Slovenia; 2 Institute J. Stefan, 

Ljubljana, Slovenia 

We present a direct measurement of the molecular velocity 

autocorrelation spectrum of bulk water by using a novel technique of 

modulated gradient spin echo method (MGSE) that reveals the effect of 

hydrogen-bonding network on the molecular self- diffusion. Structure 

and dynamics of the hydrogen-bonding network are what makes 

water unique. Understanding these phenomena has been one of the 

most important scientific challenges of the past 100 years. Although 

the long-range structure of water and its dynamics have never been 

clearly experimentally determined, numerous theoretical models 

and simulations exist, that give some explanation. Inelastic neutron 

scattering and photon scattering can probe indirectly the hydrogenbond 

dynamics at high frequencies. However, the MGSE method allows 

a direct measurement of the VAS in the frequency range determined 

by the rate of the sequence of radiofrequency and gradient pulses. 

Method can select the molecular displacements according to their time 

scales and, thus, clearly distinguishes a fast diffusion-like motion from 

a slow collective motion, like, for example, flow or convection in fluids, 

which usually introduce artifacts in other methods. The pulsed-gradient 

version of the MGSE method has the limit to the modulation frequencies 

at below 1 kHz because of the gradient coil inductance. We have 

introduced a novel technique of MGSE that enables the measurement 

at frequencies above 10 kHz. Its application to the bulk water gives an 

unusual low-frequency spectrum, similar to that of restricted diffusion 

in porous media. We explain this decrease in terms of the coupling 

between a diffusing molecule and the “flickering” attractive points, i.e., 

the broken bonds in the hydrogen-bond network. The description of the 

low-frequency dynamics in water by the harmonically coupled Langevin 

equations gives the spectrum that fits well to experimental data. In this 

way, we can determine the coupling strength of water to the attractive 

centers and the network ordering at various temperatures. The result 

confirms the direct impact of the network-like structure of water on the 

molecular self-diffusion and thus does uncover some wet secrets. 


ABSTRACTS ORAL

ABSTRACTS POSTER 

Bi10 

Optimizing incorporation of isotopically labeled amino acids 

with cell-free expression 

Pedersen, Anders; Karlsson, B. Göran 

University of Gothenburg, Swedish NMR Centre, Göteborg, Sweden 

Protein sample production for NMR using cell-free expression holds 

several advantages compared to in vivo expression techniques. Isotope 

scrambling is greatly suppressed as protein synthesis is made in a 

cellular extract of limited remaining metabolism, i.e. aminotransferase 

activities. The problems often associated with production of deuterated 

proteins by regular expression methods and the concomitant incomplete 

back-exchange of protons to the peptide amides in e.g. membrane 

proteins can be avoided by using in vitro expression as it is performed in 

regular water where only the supplied amino acids are deuterated. We 

have set up an in-house cell-free system and are currently optimizing 

the incorporation of labeled amino acids by selectively removing 

offending activities of the cellular extract that consume amino acids 

nonproductively. Activities metabolizing serine and glutamine have been 

identified and inhibited, consequently the synthesis yield has increased 

some 2-3-fold depending on which protein is expressed. 

Bi11 

40 

Biomolecules 

NMR studies of multidomain uncoupling in Syk kinase, an 

order-disorder allosteric transition regulates receptor binding 

Post, Carol Beth; Zhang, Yajie; Geahlen, Robert 

Purdue University, Med Chemistry, W Lafayette IN, United States 

The impact of tyrosine phosphorylation on Syk protein-tyrosine kinase 

conformation and functional interactions has been investigated using 

NMR structural and dynamic studies. Syk, an essential protein in 

immune signaling, binds B cell antigen receptor (BCR) with nM affinity 

but must be subsequently released to continue proper signaling in 

the cytoplasm. The high affinity binding is known to result from the 

bifunctional binding of a tandem pair of SH2 domains (tSH2) in Syk to 

a doubly-phosphorylated motif (dp-ITAM) of BCR. On the other hand, 

release from the BCR appears to result from tyrosine phosphorylation 

far from the binding sites, so that the physical basis for dissociation 

is less clear. Heteronuclear relaxation and residual dipolar couplings 

(RDCs) were used to probe the mechanism for releasing Syk from 

the BCR upon tyrosine phosphorylation. Comparison of Syk tSH2 and 

the variant tSH2pm, with the essential tyr substituted by glu, support 

an allosteric mechanism whereby release is triggered by an order 

to disorder transition in response to the negative charge of tyrosine 

phosphorylation at the distant site. Remarkable differences in relaxation 

rates and RDCs for tSH2 and tSH2pm show that phosphorylation 

partly uncouples the tSH2 domains and induces conformational 

disorder. The RDC data also establish that the unbound tSH2 solution 

structure is inconsistent with the crystallographic structure. Further, 

Syk tSH2pm has significantly lower affinity for dp ITAM (~100-fold 

weaker) than tSH2, and the impact of negative charge by tyrosine 

phosphorylation on domain orientation of these complexes is being 

investigated using RDCs. We propose that partial uncoupling, as 

opposed to complete uncoupling, of the SH2 domains is critical for 

actively inhibiting bifunctional binding and for promoting release 

when Syk phosphorylation occurs on receptor bound Syk. A model for 

the allosteric mechanism of receptor release triggered by a disorder 

transition will be presented. 

Bi12 

Structural stability modulates ligand affinity in cellular retinolbinding 

proteins types I and II 

Lücke, Christian 1 ; Cavazzini, Davide 2 ; Rossi, Gian Luigi 2 ; Franzoni, 

Lorella 3 

1 Max Planck Research Unit for Enzymology of Protein Folding, Halle 

(Saale), Germany; 2 University of Parma, Department of Biochemistry 

and Molecular Biology, Parma, Italy; 3 University of Parma, Department of 

Experimental Medicine, Parma, Italy 

Vitamin A trafficking and metabolism are regulated by specific highaffinity 

carriers. Inside the cell, this task is performed primarily by 

the cellular retinol-binding proteins types I and II (CRBP-I, CRBP-II). 

These two isoforms display a different tissue distribution and appear 

to play distinct roles in the maintenance of vitamin A homeostasis. 

They belong to a larger family of ubiquitous intracellular lipid-binding 

proteins (i-LBPs), where the hydrophobic ligand is bound within a waterfilled 

cavity inside a β-barrel structure that consists of ten antiparallel 

β-strands and is covered by two short α-helices. 

Although both CRBP types exhibit the same structural topology and 

identical binding motifs, the K d value (nM range) is about 100fold 

lower in CRBP-I with respect to CRBP-II. The molecular basis for this 

dramatic discrepancy in retinol affinity was hitherto unknown. Here we 

present recent results from NMR-based hydrogen/deuterium exchange 

experiments, which indicate that this difference in ligand affinities is 

correlated with the structural stabilities of the two proteins. The data 

revealed slower exchange of the amide protons in each holo form 

compared to the respective apo forms, which can be attributed to a 

more rigid overall protein structure due to the presence of the ligand, 

as observed also for other members of the i-LBP family. Remarkably, 

in general a slower amide proton exchange was detected for CRBP-I, 

indicating a reduced flexibility of the backbone structure relative to 

CRBP-II, both in the absence and presence of retinol. 

The significant difference in the structural stability between these two 

close protein homologues is the result of very specific amino acid 

substitutions during the course of evolution, which will be discussed 

in detail. The structural stability of these lipid carriers apparently thus 

modulates ligand uptake and release. 

Bi13 

Structural insights into the RNA-dependent RNA polymerase of 

respiratory syncytial virus 

Sizun, Christina 1 ; Bontems, François 1 ; Dubosclard, Virginie 2 ; Eleouet, 

Jean-François 2 

1 CNRS, ICSN, Gif-sur-Yvette, France; 2 INRA, VIM, Jouy-en-Josas, France 

Human Respiratory Syncytial Virus (RSV) is a major cause of severe 

pediatric respiratory tract disease worldwide. To date no vaccine is 

available for this Paramyxovirus. The RNA-dependent RNA polymerase 

(RdRp) of RSV, which recognizes the ribonucleoprotein transcription/ 

replication complex, constitutes an ideal target for antiviral drugs. 

However structure and function of the RSV RdRp complex are poorly 

understood. One of the central questions is how the polymerase 

switches between replication and transcription. 

The RdRp consists of the large polymerase (L) and a number of cofactors 

including phosphoprotein (P) and nucleoprotein (N) as well 

as two small proteins M2-1 and M2-2, that act as transcription and 

replication co-factors. Our aim is to obtain structural data of these 

proteins and to characterize their interaction domains. 

We have focussed on two co-factors, P and M2-1, and started to 

investigate them by NMR. M2-1 is an essential transcription factor of 

RSV specific to Pneumoviruses. This protein binds RNA and interacts 

with P and N proteins. It forms tetramers and is organized in four 

sub-domains: a putative Zinc finger, an oligomerization domain, a 


globular core domain that competitively interacts with P and RNA, and 

an unstructured C-terminal region. We have solved the structure of 

the core domain by NMR and started to investigate the interactions 

with RNA and P protein by chemical shift mapping. Protein P on the 

other hand forms tetramers and seems to consist mainly of disordered 

regions. Several truncated versions of P protein have been produced, 

including or not the oligomerization domain. We confirmed by NMR 

that the 10 C-terminal residues interact with the N:RNA complex, 

which forms the template recognized by the RSV RdRp. In contrast to 

P proteins of other Paramyxoviruses, the N-binding domain of RSV-P is 

not an ordered helical sub-domain. 

Bi14 

NMR structures of ubiquitin-binding motifs of translesion 

synthesis polymerase iota 

Burschowsky, Daniel 1 ; Rudolf, Fabian 2 ; Herrmann, Torsten 3 ; Peter, 

Matthias 1 ; Wider, Gerhard 1 

1 ETH Zurich, Department of Biology, Zurich, Switzerland; 2 ETH Zurich, 

Department of Biosystems Science and Engineering, Basel, Switzerland; 

3 CNRS / ENS Lyon / UCB Lyon 1, Centre Européen de RMN à Très Hauts 

Champs, Lyon, France 

Recently, a variety of novel ubiquitin-binding domains (UBDs) were 

discovered to be involved in DNA damage response; two UBDs (UBM 

and UBZ) are evolutionarily conserved in all Y-family TLS polymerases 

[1]. Translesion synthesis (TLS) is the major pathway by which 

mammalian cells replicate across DNA lesions, such as thymine dimers. 

The process is regulated by interactions with monoubiquitinated 

proliferating cell nuclear antigen (PCNA) and the UBDs of the 

polymerases, details are still unclear [2]. 

In this study we investigated the binding of two murine UBMs 

(UBM1 and UBM2) of TLS polymerase iota to ubiquitin by NMR and 

fluorescence spectroscopy. The UBMs were investigated as constructs 

of 50 amino acids in length, with the predicted 30 amino acid domains 

in the center. The solution structures of the UBM domains have been 

determined in complexes with ubiquitin rather than calculating the free 

UBM structures of the UBMs and placing them on the known ubiquitin 

structure via molecular modeling. Based on intra- and intermolecular 

NOE data of the UBM/ubiquitin complexes, we calculated de novo 

structures of the entire complexes. 

The UBMs use a different binding surface than most other UBDs that 

bind on the patch around Ile44. The UBMs’ binding surfaces center 

around Leu8 and binding isn’t abolished in Ile44Ala mutants. There are 

several surface amino acids in this region on ubiquitin that didn’t show 

any phenotypes upon substitution with alanine [3]. We investigated 

several of these ubiquitin mutants on their binding behavior with the 

UBMs by fluorescence and NMR titrations and we tested their impact on 

UV sensibility in yeast. 

1. Hofmann, K., Ubiquitin-binding domains and their role in the DNA 

damage response. DNA Repair, 2009, Epub ahead of print. 

2. Bienko, M., et al., Ubiquitin-binding domains in Y-family polymerases 

regulate translesion synthesis. Science, 2005. 310(5755): p. 1821- 

1824. 

3. Sloper-Mould, K.E., et al., Distinct functional surface regions on 

ubiquitin. Journal of Biological Chemistry, 2001. 276(32): p. 30483- 

30489. 


Bi15 

Diverse helix-helix interactions in dimeric single-span 

transmembrane proteins studied by combined use of NMR 

spectroscopy and molecular modeling 

Bocharov, Eduard; Volynsky, Pavel; Mineev, Konstantin; Nadezhdin, Kirill; 

Pustovalova, Yulia; Mayzel, Maxim; Goncharuk, Marina; Bocharova, 

Olga; Polyansky, Anton; Efremov, Roman; Arseniev, Alexander 

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Division 

of Structural Biology, Moscow, Russian Federation 

The interaction between transmembrane helices is of a great interest 

because it directly determines biological activity of a membrane protein. 

Either destroying or enhancing such interactions can result in many 

diseases related to dysfunction of different tissues in human body. 

One of the most common forms of membrane proteins is a dimer 

containing two membrane-spanning helices associating laterally to 

form a tight complex. Development of new types of drugs targeting 

membrane proteins requires precise structural information about this 

class of objects. Recent development of NMR and molecular modeling 

techniques made it possible studies of the nature and the mechanisms 

of important interactions inside the membrane complexes. To investigate 

spatial structure and internal dynamics of such transmembrane 

helical dimers in membrane mimic environment such as detergent 

micelles and lipid bicelles, we have developed a robust strategy 

using a combination of NMR spectroscopy and molecular modeling. 

The developed approach was successfully applied to recombinant 

transmembrane fragments of several proteins from different families, 

which play important roles in normal and in pathological conditions 

of human organism by providing cell signaling, maintaining cellular 

homeostasis and controlling cell fate. We characterized diverse 

transmembrane helix packing interfaces and obtained detailed atomistic 

picture of the intra- and intermolecular (protein-protein, proteinlipid 

and protein-water) interactions, that along with the available 

biochemical data provided useful insights into the protein functioning in 

norma and pathology. 

Bi16 

Structural analysis solely by NMR spectroscopy of the 

exopolysaccharide produced by Streptococcus thermophilus 

ST1 

Säwén, Elin 1 ; Huttunen, Eine 2 ; Zhang, Xue 3 ; Yang, Zhennai 3 ; Widmalm, 

Göran 1 

1 Stockholm University, Organic chemistry, Stockholm, Sweden; 

2 University of Helsinki, Department of Food Technology, Helsinki, Finland; 

3 Northeast Agricultural Research Center of China, Center of Agro-food 

Technology, Changchun, China 

Streptococcus thermophilus is a major dairy starter used in yoghurt and 

cheese production. The texture of yoghurt is strongly dependent on the 

physical and structural properties of the EPS, such as type (capsular 

or ropy), sugar composition and degree of branching. In the present 

study, the S. thermophilus strain ST1 was found to produce a viscous 

EPS when grown in skim milk. The EPS contains a hexasaccharide 

repeating unit of D-galactose and D-glucose residues, which is a new 

EPS structure of the S. thermophilus species. 

The structure of the EPS produced by S. thermophilus ST1, has been 

elucidated solely by NMR spectroscopy. The sugar composition was 

determined from 1 H and 13 C chemical shifts and coupling constant 

patterns and was confirmed by the absolute configuration determined 

by 1 H NMR spectroscopy. To elucidate the primary structure 1 H and 13C 

NMR spectroscopy, including 2D 1 H,1H TOCSY, 1 H, 1 H NOESY, 1 H, 13 C 

HSQC, 1 H,13C H2BC and 1 H, 13 C HMBC experiments were used. To 

resolve ambiguities due to spectral overlap in the 2D NOESY spectra, 

tilted projections of a three dimensional 1 H, 1 H NOESY- 1 H, 13 C HSQC 

ABSTRACTS POSTER


spectrum were recorded. By the use of tilted projections where the 13 C 

and 1 H evolutions are linked together the need to record a full threedimensional 

spectrum was avoided. 

Bi17 

Solid-state NMR analysis of peptides in membranes: influence 

of dynamics and isotope labeling scheme 

Strandberg, Erik 1 ; Esteban-Martín, Santi 2 ; Salgado, Jesús 2 ; Ulrich, Anne 

S 1 

1 Karlsruhe Institute of Technology, Institute for Biological Interfaces, 

Karlsruhe, Germany; 2 Universidad de Valencia, Instituto de Ciencia 

Molecular, Valencia, Spain 

By isotope labeling of membrane-bound peptides, typically with 2 H, 

19 F, or 15 N, solid-state NMR experiments can yield data from which 

the orientation of peptides in a native membrane environment can 

be determined. Such an orientation is defined by a tilt angle and an 

azimuthal rotation angle. 

Here we show that to obtain correct values of the orientation angles, 

dynamics should be included in the analysis of the NMR data. The 

effects of dynamics are different depending on the type of isotope 

labeling and NMR experiment considered. 

For simulated 2 H- or 19 F-NMR data, even moderate motions were found 

to have a large influence, as calculated tilt values are consistently much 

too small, unless dynamics is properly considered. A simple dynamic 

model, including a molecular order parameter scaling factor, gives good 

results only for moderately mobile peptides, while for high mobility 

cases the correct tilt is only obtained by re-introducing the explicit 

dynamics in the fitting functions. In contrast, 15 N-NMR data appear to be 

less sensitive to rigid-body peptide motions, and PISEMA spectra can 

give correct orientations even for highly mobile peptides, and assuming 

a static model for the analysis. The differences are due to the different 

orientation of the tensors of 2 H- and 19 F-labels, placed on peptide side 

chains, compared to the orientation of the 15 N tensor, placed on amide 

backbone groups. 

In particular, the tilt angles of transmembrane peptides can be severely 

underestimated from 2 H-NMR if dynamics is not taken into account. We 

show that previous 2 H-NMR experimental data of model transmembrane 

peptides in membranes of different thickness can be re-interpreted by 

explicitly considering whole-body dynamics. The result is a new set of 

tilts which follows nicely the hydrophobic mismatch expectations, and 

is coherent with molecular dynamics simulations as well as with other 

mismatch studies conducted with natural protein fragments. 

We conclude that dynamics should be included in the analysis of solidstate 

NMR data of membrane-bound peptides. Not only does this give 

more accurate orientations, but it can also provide information about the 

dynamics of the peptide. 

Bi18 

Dynamically driven ligand selectivity in cyclic nucleotide 

binding domains 

Melacini, Giuseppe 

McMaster University, Chemistry & Biochemistry, Hamilton, Canada 

One of the mechanisms that minimize the aberrant cross-talk between 

cAMP- and cGMP-dependent signalling pathways relies on the 

selectivity of cAMP-binding domains (CBDs). For instance, the CBDs of 

two critical eukaryotic cAMP receptors, i.e. protein kinase A (PKA) and 

the exchange protein activated by cAMP (EPAC), are both selectively 

activated by cAMP, but the determinants of their cAMP vs. cGMP 

selectivity are quite distinct. In PKA this selectivity is controlled primarily 

at the level of ligand affinity, whereas in EPAC it is mostly determined 

at the level of allostery. However, the molecular basis for these distinct 

42 

selectivity mechanisms is not fully understood. We have therefore 

comparatively analyzed by NMR the cGMP-bound states of the essential 

CBDs of PKA and EPAC, revealing key differences between them. 

Specifically, cGMP binds PKA preserving the same syn base orientation 

as cAMP at the price of local steric clashes which lead to a reduced 

affinity for cGMP. Unlike PKA (1,2), cGMP is recognized by EPAC in 

an anti conformation and generates several short- and long-range 

perturbations. While these effects do not alter significantly the structure 

of the EPAC CBD, remarkable differences in dynamics between the 

cAMP- and cGMP-bound states are detected, suggesting that one of 

the determinants of cGMP antagonism in EPAC is the modulation of the 

entropic control of inhibitory interactions (3). Our study illustrates the 

pivotal role of allostery in determining signalling selectivity as a function 

of dynamic changes. 

(1) Das R, et al. Taylor SS, Melacini G. Proc. N. Ac. Sci. USA 2007; 104: 

93-8; 

(2) Das R, Melacini G. J Biol Chem. 2007; 282(1):581-93; (3) Das R. et 

al. Melacini G. J. Biol. Chem. 2008, 283(28):19691-703. 

Bi19 

Comparison of the non-native states of the homologous 

proteins human and hen lysozyme and their mutants 

Wirmer-Bartoschek, Julia; Heinicke, Friedericke; Silvers, Robert; 

Schwalbe, Harald 

Goethe-Universität Frankfurt, BMRZ, Frankfurt, Germany 

The importance of protein misfolding and aggregation for human 

diseases such as Parkinson‘s, Alzheimer’s and Creutzfeldt–Jakob 

disease has been recognized in recent years. It is now well accepted 

that in particular partially unfolded or unfolded states are precursors 

of the states in the beginning of the diseases. Here, we report on the 

influence of single point mutants on the residual structure and dynamics 

of the homologous unfolded proteins hen lysozyme and human 

lysozyme. While human lysozyme mutants exist that are known to form 

fibrils in vivo, fibril formation in vivo is not known for hen lysozyme. A 

comparison of structural and dynamical properties of human and hen 

lysozyme, both in their reduced states, i.e. in the absence of any native 

disulfide bridge will be shown. Furthermore, effects of single point 

mutants of the respective proteins will be discussed. 

We have used chemical shifts and RDC measurements and in particular, 

transverse relaxation rates (R2), rotating-frame longitudinal (R1rho) 

relaxation rates and Relaxation dispersion (Rex) experiments to access 

structural and dynamical properties. 

The non-native states of human and hen lysozyme vary significantly 

despite the high sequence identity (> 60%). Changes caused by the 

amyloidogenic variants D67H and I56T will be discussed. Single point 

tryptophan to glycine mutants in hen lysozyme change hydrophobic 

clusters present in the non-native state considerably, as we can show 

consistently using RDCs and relaxation measurements. 

Furthermore, we can show that the presence and absence of single 

disulfide bridges modulates the dynamics of the model protein hen 

lysozyme investigated proteins drastically. 

The results will be discussed in the view of productive protein folding 

and in the view of amyloid formation. 


Bi20 

Influence of unfolded state dynamics and structure on fibril 

formation of human prion protein hPrP(121-230) 

Wirmer-Bartoschek, Julia; Gerum, Christian; Silvers, Robert; Schwalbe, 

Harald 

Goethe-Universität Frankfurt, BMRZ, Frankfurt, Germany 

Transmissible spongiform encephalopathies are characterized by 

accumulation of the abnormal “scrapie” form (PrPSc) of the endogenous 

cellular form (PrPC) of the prion protein (PrP) in the brain. The 

conversion of soluble PrPC to the pathogenic form involves largescale 

rearrangement of tertiary structure to convert the native benign 

state of the protein to a highly order fibril aggregate. It is commonly 

accepted that unfolded intermediates are significantly populated 

during this conversion process but the detailed mechanism of this 

conversion including the influence of disease-related mutations 

remains unknown. Here, we investigate the unfolded state by NMR 

spectroscopy to characterize structure and dynamics of the third 

important conformational state of the prion protein. We show that 

the loss of the native disulfide bridge (Cys179–Cys214) changes 

its structural dynamics. The unfolded state of hPrP(121–230) has 

considerable degree of residual structure with beta-state propensities 

both in the oxidised and in the reduced form. However, as evidenced by 

15N R2 and R1rho relaxation rates, the formation of the native disulfide 

(Cys179–Cys214) in the oxidised state rigidifies the surrounding amino 

acids (Phe175–Glu221), a hotspot region of the protein regarding 

disease-related mutations that promote aggregation. The change in 

the structural dynamics has a pronounced effect on the aggregation 

behaviour of the protein, only the oxidised protein forms fibrils. 

Bi21 

Effects of dynamics in 4D protein 1 H chemical shift prediction 

Lehtivarjo, Juuso 1 ; Hassinen, Tommi 1 ; Korhonen, Samuli-Petrus 2 ; 

Peräkylä, Mikael 1 ; Laatikainen, Reino 1 

1 University of Kuopio, Department of Biosciences, Kuopio, Finland; 

2 Perch Solutions Ltd., Kuopio, Finland 

An empirical method for protein 1 H chemical shift prediction, called 

4DSPOT, is developed [1]. Since proteins are flexible structures in 

liquid phase, their motions were included in the prediction method as 

a ‘fourth’ dimension. A database consisting 40 protein models (> 21 

000 1 H chemical shifts) was built up. To model protein motions in liquid 

phase, 150 ps molecular dynamics simulations were performed for 

each protein model with AMBER 9 program [2]. The predictor program 

builds a number of time-averaged molecular descriptors, such as 

torsion angle, anisotropy and solvation terms, to describe the chemical 

environment of each proton. Prediction algorithm is mathematically 

based on principal component analysis (PCA). 

In this presentation, effects of protein dynamics to prediction results are 

investigated. For the set of 40 proteins, some remarkable improvements 

in prediction accuracy were observed. Only in few cases, the use of 4D 

prediction deteriorated the results. Averagely, 4D prediction decreases 

the RMS errors by 7 and 6 % for Hα and HN nuclei, respectively. 

[1] Lehtivarjo J; Hassinen T; Korhonen S-P; Peräkylä M; Laatikainen, R. 

To be submitted to J. Biomol. NMR (2009). 

[2] Case D.A; Cheatham, T.E. III; Darden T; Gohlke H; Luo R; Merz, K.M; 

Onufriev A; Simmerling C; Wang B; Woods R. J. Computat. Chem. 26, 

1668-1688 (2005). 


Bi22 

Lipid composition of the bacterium Helicobacter pylori analyzed 

using a new 31 P, 1 H correlation experiment 

Petzold, Katja 1 ; Olofsson, Annelie 1 ; Johansson, Pär 1 ; Arnqvist, Anna 1 ; 

Gröbner, Gerhard 2 ; Schleucher, Jürgen 1 

1 Umeå University, Dept. of Medical Biochemistry and Biophysics, Umeå, 

Sweden; 2 Umeå University, Department of Chemistry, Umeå, Sweden 

The gram-negative bacterium H.pylori is carried by over half of 

the human population and causes peptic ulcer disease and gastric 

cancer. H.pylori lives under harsh gastric conditions (pH 1-4), where 

persistent infection relies on bacterial adherence to the gastric mucosa, 

mediated by membrane proteins such as Lewis B binding adhesin, 

BabA. Furthermore the bacterium sheds vesicles to deliver bacterial 

virulence factors. Adhesion and shedding of vesicles require a functional 

membrane environment; therefore we analyze H.pylori membranes to 

find connections between membrane composition, adhesion activity of 

membrane proteins, and virulence. 

For this purpose, we developed a semi-constant time 31 P, 1 H COSY with 

gradient coherence selection, to unambiguously identify and quantify 

phospholipids. The semi-constant time evolution of 31 P achieves high 

resolution in the 31 P dimension and comparable transfer efficiency for 

different lipids allow quantification. In the 1 H dimension, 31 P, 1 H and 

1 H, 1 H J-couplings are used to unambiguously assign phospholipids, 

independent of changes in chemical shifts. Lipid composition can be 

determined semi-quantitatively by integrating the 31 P projection of the 

2D spectrum. 

We analyzed lipids from whole cells, inner- (IM) and outer membranes 

(OM), and vesicles from H.pylori and detected variation in phospholipid 

composition depending on growth; interestingly this change was 

correlated to activity of the BabA adhesin. Lipids of vesicles are closely 

related to the OM and differ from IM, so that we conclude that they 

are derived from the OM. Contrary to expectation, the concentration of 

Cardiolipin, known to enhance membrane curvature, is not related to 

the size of vesicles. The sensitive 31 P, 1 H COSY experiment can address 

how lipids are involved in the control of H.pylori virulence, and may be 

useful for other classes of organic Phosphor compounds. 

Bi23 

Distinct targeting of DnaK with novel peptidic antibiotics 

Heydenreich, Katja; Liebscher, Markus; Chao, Yu; Schiene-Fischer, 

Cordelia; Lücke, Christian 

Max Planck Research Unit for Enzymology of Protein Folding, Halle/ 

Saale, Germany 

The prokaryotic HSP70 analogue DnaK acts as a holding chaperone 

and a cis/trans isomerase in assisted protein folding processes. DnaK is 

essential for cell division and bacterial viability under stress conditions. 

Highly specific and potent DnaK inhibitors are therefore advantageous 

for the design of effective antibacterial agents. 

Based on a peptide library, screening the N-terminal part of the 

antimicrobial peptide pyrrhocoricin, a number of derivatives were 

developed in our lab that either display an improved binding affinity to 

DnaK or inhibit peptidyl cis/trans isomerase activity and DnaK/DnaJ/ 

GrpE-assisted refolding of firefly luciferase – or in some cases even 

both. 

Here we characterize by chemical shift perturbation the DnaK-binding 

properties of several inhibitors as well as a substrate peptide. The 

employed pyrrhocoricin-like peptides differed in amino acid sequence, 

peptide length and membrane permeability. We used the construct 

DnaK 387-552 , which represents the substrate-binding domain of DnaK. 

Sequential 1 H, 13 C and 15 N resonance assignments were performed by 

means of three-dimensional HNCA and HNCACB experiments. 

In agreement with X-ray data, our NMR results demonstrate a 

ABSTRACTS POSTER


comparable binding mode for all peptides, generally involving a central 

leucine side-chain that is buried in a hydrophobic cavity of DnaK. 

Furthermore the data indicate that the newly developed high-affinity 

peptide derivatives fill the relatively spacious funnel on the DnaK 

surface rather compactly, thus achieving a more specific binding to the 

substrate-binding site of the protein. 

Bi24 

Structural investigations of dimeric transmembrane domain of 

amyloid precursor protein 

Nadezhdin, Kirill; Bocharova, Olga; Bocharov, Eduard; Arseniev, 

Alexander 

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Division 

of Structural Biology, Moscow, Russian Federation 

The amyloid precursor protein (APP) undergoes sequential cleavages 

to generate diverse polypeptides, including the amyloid beta peptide 

(Abeta), which is believed to play a major role in amyloid plaque 

formation in Alzheimer’s disease. However, its physiological functions 

remain unclear. It is well known that specific familial mutations in APP 

cause overproduction of Abeta. The majority of these defects is located 

in the transmembrane (TM) domain of APP, probably disordering the 

protein fold and influencing its dimerization mechanism in the plasma 

membrane. The presence of characteristic sequential motifs in the APP 

polypeptide chain, such as GXXXG/A, indicates the helix-helix type of 

TM dimerization. We developed the bacterial expression system and 

purified recombinant APP fragment (686-726) including the TM domain. 

Using the modern solution NMR techniques we investigated structuraldynamic 

properties of the dimeric APP TM domain in membrane 

mimetic environment. 

Bi25 

Spatial structures of membrane domains of ErbB receptor 

kinases 

Mineev, Konstantin 1 ; Bocharov, Eduard 1 ; Pustovalova, Yulia 1 ; Volynsky, 

Pavel 2 ; Tkach, Elena 3 ; Arseniev, Alexander 1 ; Efremov, Roman 2 

1 IBCh RAS, Biomolecular NMR Spectroscopy, Moscow, Russian 

Federation; 2 IBCh RAS, Biomolecular Modeling, Moscow, Russian 

Federation; 3 IBCh RAS, Protein Engineering, Moscow, Russian 

Federation 

Membrane proteins are now of the great interest in biology and 

medicine, but their spatial structures are still studied poorly. X-ray 

experiences problems with crystallization and NMR cannot deal with 

high weight and low mobility of protein-lipid complexes. Taking into 

account that membrane complexes are the most popular targets for 

modern drugs, we can postulate that understanding of principles, 

determining organization, folding and function of channels and 

receptors, is necessary. 

As the major part of membrane proteins consists of several membranespanning 

helices, in order to study such principles, we have chosen the 

simplest and the smallest object among all membrane proteins - helical 

dimers. For the calculation of protein structure special protocol was 

developed, employing 13C-filtered NOESY techniques for the detection 

of intermolecular interactions; special peak list calibration procedure 

and multiple (ambiguous) distance restraints for proper handling of 

NOE data gathered from spectra with overlapped signals with different 

dynamical characteristics. 

Using the protocol, structures of several dimers of transmembrane 

segments of ErbB-family kinase receptors were calculated: ErbB2, 

ErbB4, ErbB1/ErbB2. All of them are parallel right-handed dimers with 

the angle between the axes of helices ranging between 40 and 50 

degrees. Helices interact trough different variants of G3G motifs and 

44 

intermolecular hydrogen bonds formed by Ser and Thr sidechains. 

Experiments with ErbB4 showed that helix-helix interactions can cause 

small changes in secondary structure of transmembrane spans, in 

donor-acceptor distance of hydrogen bonds, for instance. 

This research together with biochemical data proved that membrane 

domain can play significant role in the function of membrane proteins, 

and gave additional evidences in the favor of rotational coupling 

activation mechanism, suggested for ErbB family. Moreover, spatial 

structures can explain the effects of some mutations in transmembrane 

helix of ErbB2 which are thought to be cancer-related. Further studies 

could allow designing drugs which target not extracellular but also 

membrane domains with high specificity. 

Bi26 

NMR structural characterization of the 6 M urea-unfolded 

ensemble of an ultrafast folding protein 

Kuhn, Lars T. 1 ; Richter, Christian 2 ; Saxena, Krishna 2 ; Schwalbe, Harald 2 ; 

Hore, P. J. 1 

1 University of Oxford, Physical & Theoretical Chemistry Laboratory, 

Oxford, United Kingdom; 2 JWG University, Center for Biological Magnetic 

Resonance, Frankfurt, Germany 

Even though 3D structural information is readily available for the 

native states of proteins, experimentally determined residue-specific 

information on kinetic intermediates and, in particular, on the unfolded 

states of proteins remains relatively sparse due to their conformational 

heterogeneity and dynamic nature. The “Trp-Cage” molecule (TC5b) 

is a small, 20-residue mini-protein which exhibits a well-defined 

hydrophobic core together with pronounced secondary and tertiary 

interactions in its native state. Also, TC5b is known to have one of 

the fastest kinetic refolding rates ever observed and, because of 

its small size, has been adopted as a benchmark for computational 

folding studies and molecular dynamics (MD) simulations. Following 

the recent observation of hydrophobic cluster formation in the 6 

M urea-denatured state of TC5b obtained from photo-CIDNP NOE 

pulse-labelling experiments, we have further elucidated the structural 

properties of the unfolded state of this highly unusual biomolecule using 

multidimensional NMR and heteronuclear backbone relaxation studies 

on a uniformly 15 N-labelled TC5b construct. Even though residual 

elements of secondary structure seem to be absent in unfolded TC5b, 

our results permit a clear identification of sequence-remote contact 

interactions between aliphatic and aromatic 1 H nuclei for those regions 

of the peptide whose amino acid side chains also exhibit significant 

cross-polarization in the pulse-labelling experiment. Moreover, an 

NOE-restrained calculation of an ensemble of unfolded TC5b molecules 

makes the direct assignment of aromatic tryptophan side chain 1 H 

nuclei as individual sources of cross-polarization feasible thereby 

further clarifying the CIDNP NOE data. Our results corroborate previous 

findings of a pre-existing hydrophobic cluster in the unfolded state of 

TC5b comprising both native and non-native contact interactions. These 

interactions appear to be crucial prerequisites for the ultrafast folding 

kinetics of the peptide as suggested both by MD simulation studies as 

well as independently performed tryptophan fluorescence quenching 

experiments on the unfolded ensembles of TC5b. 


Bi27 

Insight into the ubiquitin-activation process. The high resolution 

3D structure of catalytic cysteine domain of mouse ubiquitinactivating 

E1 enzyme studied by NMR spectroscopy 

Jaremko, Lukasz 1 ; Jaremko, Mariusz 2 ; Zhukov, Igor 2 ; Bochtler, 

Matthias 3 ; Szczepanowski, Roman H. 3 ; Filipek, Renata 3 ; Wojciechowski, 

Marek 3 

1 Institute of Biochemistry and Biophysics / Warsaw University, 

Labolatory of Biological NMR / MISDoMP, Warsaw, Poland; 2 Institute of 

Biochemistry and Biophysics, Labolatory of Biological NMR, Warsaw, 

Poland; 3 International Institute of Molecular and Cell Biology, Warsaw, 

Poland 

Ubiquitin-activating enzyme E1, is the first enzyme in the ubiquitinylation 

cascade which converts ubiquitin to the adenylate and subsequently to 

an enzyme-linked thioester in an ATP-dependent process. It was proved, 

that the E1 consists of “adenylation” and “catalytic cysteine” domains. 

Our previous works has shown that this domain can be divided into two 

subdomains – “first” and “second” catalytic cysteine half-domain, FCCH 

and SCCH, respectively – that fold autonomously. Recently, we reported 

the 1H, 13C, and 15N chemical shifts for the recombinant FCCH halfdomain 

of mouse E1 enzyme. The three-dimensional structure of FCCH 

half-domain was also solved by means of NMR spectroscopy. The 

ubiquitin E1 FCCH domain differs significantly from the corresponding 

FCCH domains from the SUMO and NEDD8 E1’s. The SCCH and FCCH 

are located in the different parts of the sequence, and they are far away 

from each other (~ 40 Å) in the 3D X-ray structure of the whole E1 

complex. The detailed inspection of cross peaks on 2D 1H-15N HSQC 

spectrum of 13C,15N-double labeled FCCH domain in the presence 

of equimolar ratio of unlabeled free SCCH exhibits the interactions 

between those two half-domains in the solution. The 13C,15N-double 

labeled SCCH half-domain (276 a.a. long, 31.3 kDa) shows a good 

dispersion of cross peaks on 2D 1H-15N HSQC spectrum. In this 

experiment around 240 1H-15N cross-peaks out of 257 expected were 

detected what makes the sequential assignment possible in the nearest 

future. Further we plan to complete backbone sequence-specific 

assignment of the SCCH half domain and perform NMR experiments 

to get structural information about whole catalytic cysteine domain of 

mouse E1 enzyme. The acquired data sets give us the possibility to 

compare 3D structures of SCCH and FCCH half domains in solution 

with crystal structure and supply the knowledge about FCCH – SCCH 

interaction and molecular dynamics processes in the catalytic cysteine 

domain. This work will help us to understand the essential catalytic 

activity of the ubiquitin-activating enzyme E1. 

Bi28 

The NMR solution structure of ghrelin and several of its 

fragments in membrane mimetic agents 

Venturi, Chiara 1 ; Jiménez-Barbero, Jesús 1 ; Casanueva, Felipe F. 2 ; Pazos, 

Yolanda 2 ; Martín-Pastor, Manuel 3 ; Díaz Hernández, María Dolores 1 

1 Centro Investigaciones Biológicas CSIC, Biología Estructural de 

Proteínas, Madrid, Spain; 2 Laboratorio de Endocrinología Molecular 

y Instituto de Investigación Sanitaria, Laboratorio de Endocrinología 

Molecular y Celular, Santiago de Compostela, Spain; 3 Santiago de 

Compostela University, Unidade de Resonancia Magnética, RIAIDT, 

Santiago de Compostela, Spain 

Ghrelin is a peptide hormone of 28 amino acids. Among other effects, 

ghrelin stimulates appetite, growth hormone production and decreases 

blood pressure. Its precursor preproghrelin is generated by the 

endoprotease prohormone convertase 1/3. Preproghrelin also encodes 

obestatin, a 23 amino acid peptide which is considered antagonist 

of ghrelin because of its inhibitory effect on feeding [1]. For ghrelin 

to be active and bind to the growth hormone secretagogue receptor 

(GSHR-1a), acylation at a serine residue (Ser3) is required. Acylated and 

desacylated ghrelins share some biological functions but have also their 

own roles, suggesting the possible existence of additional receptors. 

The primary sequence of ghrelin suggests a possible formation of a 

short α-helix in the central segment of the peptide but the latter has not 

been experimentally demonstrated. Theoretical studies (MD) performed 

on ghrelin suggest a short α-helix [2]. However, NMR and CD studies in 

solution (pH 1.0-1.5) revealed random coil structure. [3] Only recently, 

a partial helix (


are presented. 

The NMR measurements revealed the detailed structures of the 

complexes and confirmed the {NH 2 , 2×N Im } binding mode in the 

physiological pH. These results clearly showed that replacing of Cys- 

S-S-Cys bridge with His-M(II)-His motif is possible in physiological 

conditions, what can be useful in designing new compounds working 

efficiently in different cell environments independently on pH. 

Bi30 

Salt bridges in protein G are present in the crystal but not in 

solution 

Williamson, Mike 1 ; Tomlinson, Jenny 1 ; Hansen, Poul Erik 2 ; Ullah, Saif 2 

1 University of Sheffield, Molecular Biology and Biotechnology, Sheffield, 

United Kingdom; 2 Roskilde University, Science, Systems and Models, 

Roskilde, Denmark 

NMR investigations have been carried out on the B1 domain of protein 

G. This protein has six lysine residues, of which three are consistently 

found to form surface-exposed salt bridges in crystal structures, while 

the other three are not. The Nζ and Hζ chemical shifts of all six lysines 

are similar and are not affected significantly by pH titration of the 

carboxylate groups in the protein, except for a relatively small titration 

of K39 Nζ. Deuterium isotope effects on nitrogen and proton are of 

the size expected for a simple hydrated amine (a result supported by 

density functional theory calculations), and also do not titrate with the 

carboxylates. The lineshapes of the J-coupled 15N signals suggest 

+ rapid internal reorientation of all NH groups. pKa values have been 

3 

measured for all charged sidechains except Glu50 and do not show 

the perturbations expected for salt bridge formation, except that E35 

has a Hill coefficient of 0.84. The main differential effect seen is that 

the lysines that are involved in salt bridges in the crystal display faster 

exchange of the amine protons with the solvent, an effect attributed 

to general base catalysis by the carboxylates. This explanation is 

supported by varying buffer composition, which demonstrates reduced 

electrostatic shielding at low concentration. In conclusion, the study 

demonstrates that the six surface-exposed lysines in protein G are 

not involved in significant salt bridge interactions, even though such 

interactions are found consistently in crystal structures. However, the 

intrahelical E35-K39 (i, i+4) interaction is partially present. 

Bi31 

Intermediate rate (ns to µs) fluctuations in barnase identified 

using pressure 

Williamson, Mike 1 ; Wilton, David 1 ; Pandya, Maya 1 ; Kitahara, Ryo 2 ; 

Akasaka, Kazuyuki 3 

1 University of Sheffield, Molecular Biology and Biotechnology, Sheffield, 

United Kingdom; 2 Ritsumeikan University, College of Pharmaceutical 

Sciences, Kusatsu, Japan; 3 Kinki University, Biotechnological Science, 

Kinokawa, Japan 

We have measured 1 H NMR chemical shifts for the ribonuclease 

barnase at pressures from 3 MPa to 200 MPa, both free and bound 

to d(CGAC). Shift changes with pressure were used as restraints to 

determine the change in structure with pressure. Free barnase is 

compressed by about 0.7%. The largest changes are on the ligandbinding 

face close to Lys-27, which is the recognition site for the 

cleaved phosphate bond. This part of the protein also contains the 

buried water molecules. In the presence of d(CGAC), the compressibility 

is reduced by approximately 70%, and the region of structural change 

is altered: the ligand-binding face is now almost incompressible 

while changes occur at the opposite face. Because compressibility 

is proportional to mean square volume fluctuation, we conclude 

that in free barnase, volume fluctuation is largest close to the active 

46 

site, but that when the inhibitor is bound, fluctuations become much 

smaller and are located mainly on the opposite face. The timescale of 

the fluctuations is between ns and µs, consistent with the degree of 

ordering required for the fluctuations, which are intermediate between 

rapid uncorrelated sidechain dynamics and slow conformational 

transitions. The fluctuations are in different regions from those identified 

on the µs-ms timescale using relaxation dispersion, demonstrating 

how motions are channeled from rapid thermal motion into slower 

functionally important motions. The high-pressure technique is 

therefore useful for characterizing motions in this relatively inaccessible 

timescale. 

Bi32 

Imperfect binding of let-7 miRNA to lin-41 mRNA studied by 

NMR 

Cevec, Mirko; Plavec, Janez 

National Institute of Chemistry, Slovenian NMR Centre, Ljubljana, 

Slovenia 

MicroRNAs (miRNAs) are small noncoding RNAs, which regulate 

developmental timing, differentiation, proliferation and apoptosis in 

various organisms. They are assembled into the appropriate RNAinduced 

silencing complex (RISC) according to their intrinsic structures. 

miRNAs then induce translational repression by binding to partially 

complementary sites on target mRNAs or direct cleavage of target 

mRNAs after perfect base pairing. 

We focused on let-7 miRNAs from nematode C. elegans. They 

regulate the expression of lin-41 mRNA after imperfect binding to 

their 3’-untranslated region. Members of let-7 family miRNA were 

isolated in many organisms. For example, in human they function as 

tumor suppressors. Biotech companies are now using let-7 miRNAs as 

targets to treat lung cancer caused by Ras oncogene. The interaction 

between let-7 miRNA and lin-41 mRNA is dependent on two conserved 

let-7 complementary sites (LCS1 and LCS2). Mutations showed that 

the nature of base-pairing, asymmetric internal loops and bulges are 

important for regulation of gene expression by miRNA. 

We prepared isotopically-labeled RNA hairpin molecules and used them 

as model complexes between let-7 miRNA and the binding sites on lin- 

41 mRNA. The 3D structures of RNA constructs were determined with 

the use of standard 2D and 3D NMR techniques. Phages were used to 

align the RNA constructs and to acquire residual dipolar couplings. NMR 

restraint computer simulations showed that molecules fold into stable 

structures consisting of two stem regions separated by an asymmetric 

internal loop. Stems were stabilized by Watson-Crick base pairs. The 

asymmetric internal loop from second binding site (LCS2) adopted a 

well defined structure. Three uracils formed a base triple, while the 

two adenines formed AA base pair. The asymmetric internal loop from 

first binding site (LCS1) was less defined. These interesting structural 

elements make complex between miRNA and mRNA different in 

comparison to a common A-form RNA. 


Bi33 

Structure of Munc13-1458-492 /Ca2+ -CaM reveals a flexible 

4 

modular 1-5-8-26 recognition motif that suits short-term 

plasticity 

Maestre-Martinez, Mitcheell1 ; Rodriguez-Castaneda, Fernando1 ; 

Coudevylle, Nicolas1 ; Becker, Stefan1 ; Brose, Nils2 ; Carlomagno, Teresa1 ; 

Griesinger, Christian1 1 2 MPI for Biophysical Chemistry, Göttingen, Germany; MPI for 

Experimental Medicine, Göttingen, Germany 

The priming process is essential for the post-docking/pre-fusion 

maturation of vesicles in regulated exocytosis [1]. The interactions 

between Munc13s and calmodulin (CaM) regulate the synaptic vesicle 

priming and synaptic efficacy in response to the residual Ca2+ signal, 

thereby shaping the short-term plasticity during periods of synaptic 

activity [2]. It was also shown that the Munc13s C1 domain lowers 

the energy barrier for synaptic vesicle fusion [3]. Munc13-1 and 

ubMun13-2 feature CaM binding sites located N-terminal to their 

C1 domains, which bind CaM in a Ca2+ -dependent manner [2]. We 

present the NMR solution structure of the Munc13-1458-492 /Ca2+ 4-CaM complex, which reveals a novel (1-5-8-26) CaM binding motif featuring 

two modules connected by flexible linkers. The C-module consists of 

an amphiphilic helix from Munc13-1 bound to the C-terminal CaM 

domain, with hydrophobic residues at positions 1-5-8 (i.e. W464, 

F468 and V471) acting as anchors, while the N-module displays a 

Trp residue located in position 26 (W489) attached to the N-terminal 

CaM domain. The structure is extended and flexible around the linkers 

connecting C- and N-module, as revealed by NOE, RDC and PCS data. 

EGTA titrations followed by HSQC spectra also revealed the existence 

of a half Ca2+ -loaded Munc13-1458-492 /Ca2+ -CaM complex, which can 

2 

interconvert rapidly into the fully loaded species. This leads to a fast 

equilibrium that allows the complex to act as an efficient Ca 2+ -sensor at 

high calcium concentrations. The same behavior was found in Munc13- 

2, which shares the same CaM recognition motif. Our results provide an 

explanation for the roles of CaM and Munc13s in short term synaptic 

plasticity. A preliminary study of the interactions of CaM with the C1 

domain of Munc13-1 is also presented. 

References: 

[1] Brose, N.; Rosenmund, C. and Rettig, J., Curr Opin Neurobiol, 2000, 

10, 303-311. 

[2] Junge, H.J.; Rhee, J-S.; Jahn, O.; Varoqueaux, F.; Spiess, J.; 

Waxham, M.N.; Rosenmund, C. and Brose, N., Cell, 2004, 118, 389- 

401. 

[3] Basu, J.; Betz, A.; Brose, N. and Rosenmund, C., J Neurosci, 2007, 

27, 1200-1210. 

Bi34 

NMR study of model systems for transaldimination in pyridoxal- 

5’-phosphate dependent enzymes 

Chan Huot, Monique; Tolstoy, Peter; Limbach, Hans Heinrich 

Freie Universität Berlin, Chemistry and Biochemistry, Berlin, Germany 

Pyridoxal-5’-phosphate (PLP) is a cofactor in many enzymes involved 

in amino acids transformations. PLP is bound covalently through an 

imine bond with a lysine side chain residue in the active site forming 

an internal aldimine. We have previously studied the protonation states 

of PLP aldimines showing the coupled protonation of the ring nitrogen 

to the phenolate in the PLP moiety of the aldimine. [1] The internal 

aldimine is the starting point of PLP dependent enzyme’s catalytic 

cycle. The first step consists in the transformation from the internal 

aldimine to the external aldimine formed between the PLP cofactor 

and the incoming amino acid. This is called the transaldimination. In 

order to study this crucial step, PLP was selectively enriched at the 

position C-4’ in order to study its interaction with model sytem of the 

internal and external aldimine. 13 C and 15 N NMR studies have shown 

that the transaldimination lies over two major conditions. The first one 

consists in considering that the dielectric constant inside the active 

site is smaller than water. [2] This involves that the amino acid enters 

the active site in a neutral form offering a nucleophilic amine to the 

sytem. The second point is the fact that the side chain of the lysine 

covalently linked to PLP prefers to be protonated and will then offer an 

electrophilic center to the incoming amine. The system will favor the 

protonated free lysine which is the driving force of the reaction. 

[1]Golubevet al., J. Mol. Struct. 2007, 844-845, 319-327 

[2] Sharif et al., J. Am. Chem. Soc. 2007, 129, 9558-9559 


Bi35 

Group epitope mapping considering relaxation of the ligand 

(GEM-CRL) 

Kemper, Sebastian 1 ; Patel, Mitul 2 ; Davis, Benjamin G. 2 ; Jones, Jonathan 

A. 3 ; Claridge, Timothy D. W. 2 

1 University of Cologne, Department of Chemistry, Cologne, Germany; 

2 University of Oxford, Chemistry Research Laboratory, Oxford, United 

Kingdom; 3 University of Oxford, Centre of Quantum Computation, 

Oxford, United Kingdom 

The saturation transfer difference (STD) 1 experiment has proven to be 

a popular technique for the investigation of protein-ligand interactions, 

because of its relative ease of application and its suitability to study 

unlabeled and large proteins. Aside from these advantages, one 

problem of the method exists in the quantitative analysis of the 

experimentally observed STD factors, in which the relaxation of the 

ligand is one of the major influences, making interpretation of these 

difficult when attempting to define a group epitope map (GEM) for the 

bound ligand. 2,3 

It will be shown that an approximation of the relaxation matrix leads 

to a simple equation in which relaxation of the ligand is accounted 

for implicitly by inclusion of its experimentally determined longitudinal 

relaxation rates (T1 time constants). In this “group epitope mapping 

considering relaxation of the ligand” (GEM-CRL), the result reflects the 

directly transferred magnetization rates from the protein onto the ligand. 

The conditions under which this approach can be applied were tested 

on a theoretical model system and a protein-saccharide complex with 

known crystal structure. These results were also compared with values 

obtained from full relaxation matrix calculations (CORCEMA-ST) 4,5 , from 

which it could be shown that the GEM-CRL methodology is superior to 

the standard group epitope mapping. 

(1) Mayer, M.; Meyer, B. Angewandte Chemie International Edition 1999, 

38, 1784-1788. 

(2) Yan, J.; Kline, A. D.; Mo, H.; Shapiro, M. J.; Zartler, E. R. Journal of 

Magnetic Resonance 2003, 163, 270-276. 

(3) Mayer, M.; Meyer, B. Journal of the American Chemical Society 

2001, 123, 6108-6117. 

(4) Rama Krishna, N.; Jayalakshmi, V. Progress in Nuclear Magnetic 

Resonance Spectroscopy 2006, 49, 1-25. 

(5) Jayalakshmi, V.; Krishna, N. R. Journal of Magnetic Resonance 

2002, 155, 106-118. 

ABSTRACTS POSTER


Bi36 

Structural basis for the differential membrane-binding of the 

oxidized and reduced TOR FATC domain 

Dames, Sonja A.; Rathgeb-Szabo, Klara; Grzesiek, Stephan 

Biozentrum, University Basel, Structural Biology, Basel, Switzerland 

The target of rapamycin (TOR) is a highly conserved ser/thr kinase 

that regulates cell growth in response to nutrient availability and 

environmental factors. The C-terminal FATC domain plays and important 

role for the regulation of the neighboring catalytic domain. It is ~35 

residues long and contains two conserved cysteines. Recently, we 

determined the structure of the oxidized form and showed by in vivo 

mutagenesis that the stability of TOR depends on the redox state of this 

domain1. Because the most C-terminal sequence stretch GWCPFW 

was suggested to represent a lipid-binding motif, we analyzed further 

the interaction of the yeast TOR1 FATC domain (= y1fatc) with different 

lipids. Initial NMR titration studies showed that oxidized y1fatc could 

bind to all tested lipids above the critical micelle concentration (CMC). 

Therefore, we determined the NMR structure of the oxidized form 

bound to dodecylphosphocholine (DPC) micelles and characterized 

its dynamic behavior by 15N relaxation experiments. The solvent 

accessibility for each residue was determined based on a titration with 

the paramagnetic ion Mn2+, the observation of NOEs to water, and the 

initial titration with DPC. 

A comparison of the 1H-15N HSQC spectra of the reduced and oxidized 

forms bound to DPC micelles revealed significant spectral differences. 

Based on a comparison of the also determined structure and dynamics 

of the reduced form bound to DPC micelles with those for the oxidized 

form, the influence of the redox-state on the membrane-binding 

properties will be discussed. In addition, we analyzed the membranceassociation 

of two trp to ala mutants. 

Membrane-binding of the FATC domain might protect it from typical 

cellular redox regulators and regulate the association with different 

cellular membranes and membrane proteins. Thereby, the presented 

data provide first molecular insights in the mechanism of TOR 

membrane-association. 

1 S. Dames et al., J. Biol. Chem, 280(21): 20558-20564, 2005 

Bi37 

Structure and backbone dynamics of Syndesmos, a new class 

of syndecan 4 interacting protein by NMR spectroscopy 

Ko, Sunggeon 1 ; Yu, Jiho 2 ; Chi, Chunhwa 1 ; Kim, Sangyun 1 ; Cho, Hyunsoo 

2 ; Lee, Weontae 1 

1 Yonsei university, Biochemistry, Seoul, Republic of Korea; 2 Yonsei 

university, Biology, Seoul, Republic of Korea 

Syndesmos, one of the syndecan 4 interacting proteins in in vitro and in 

vivo, is expressed in several organs. When syndesmos is overexpressed 

in the cell, cell spreading and cytoskeleton organization was improved 

which was confirmed by enhanced formation of filipodia suggesting 

that cellular features by syndesmos overexpression is related with 

interaction of syndecan 4 containing cell adhesion complex. Syndecan 

protein family consisting of syndecan 1, 2, 3 and 4 is the one of the 

famous single transmembrane protein for cell adhesion which is the 

heparin sulfate proteoglycan. Syndecan 4 is expressed in wide scope 

of the cells and interacts with cytoskeleton and controls the activity 

of protein kinase C (PKC ). The cytoplasmic domain of syndecan 4 

consists of three individual regions, conserved 1 region (C1 region), 

variable region (V region) and conserved 2 region (C2 region). Each 

region interacts with individuals which C2 region interacts with PDZ 

domain proteins and V region interacts with phosphatidyl inotitol 4, 

5-bisphosphate (PIP2) and syndesmos. Recently, it was reported that 

syndesmos interacts with the V region of syndecan 4 and participated 

on focal adhesion complex formation suggesting that the cellular 

48 

function of syndesmos is related with syndecan 4 and focal adhesion 

directly. Interestingly, syndesmos showed high amino acid sequence 

homology with nudix hydrolase which is the ubiquitous enzyme for 

pyrophosphate hydrolysis. The DNA sequence and amino acid sequence 

similarity between syndesmos and nudix hydrolase implied that 

syndesmos is a paralogous protein and gene duplication occurred in 

tetrapod lineage near the amniote divergence for nudix hydrolase. To 

identify the structure and function of syndesmos, we executed NMR 

spectroscopy, backbone dynamics together with X-ray crystallography 

and surface plasmon resonance (SPR) experiments. Our findings 

suggest that the structure of syndesmos shows nudix fold consisted of 

7 β-strands and 7 α-helixes and it interacts with cytoplasmic domain of 

syndecan 4 though the C1 and V region of syndecan4. 

Bi38 

NMR characterization of the near native energy landscape of 

SUMO from drosophila melanogaster (dSmt3) 

Singh, Venus; Kumar, Dinesh; Hosur, Ramakrishna V 

Tata Institute of Fundamental Research, Chemical Sciences, Mumbai, 

India 

The structure-function paradigm claims that a specific function of a 

protein is determined by its unique and rigid three-dimensional (3D) 

structure. However, proteins undergo various conformational fluctuations 

while doing their job. This suggests that the native structure of protein is 

not a well defined single state but is an ensemble of the dynamic states 

inter-converting at ms to µs time scale. These ensembles of states are 

termed as near-native states (i.e. all the structural units remain intact). 

Characterization of near-native excited states of a protein provides 

insights into various biological functions such as co-operativity, ligand 

binding/release, and protein-protein interactions. Such states can be 

populated by mild perturbations such as addition of low concentration 

of chemical denaturants, or small changes in temperature, pressure, pH 

etc. These structural perturbations get reflected in NMR chemical shift 

changes of the individual residues. 

In this work, we investigated the native energy landscape of SUMO from 

drosophila melanogaster (dSmt3). SUMO family proteins are involved 

in post-translational modification of various cellular proteins. The major 

goal is to understand the relationship between the structures (and 

dynamics) of SUMO proteins and their biological functions. Here, the 

temperature dependence of amide proton chemical shifts are used 

to characterize the residues in dSmt3 which can access alternative 

conformations. We noticed many interesting patterns as the urea 

concentration was changed from 0 M to 1.0 M (within the near native 

state region). More and more residues showed curved profiles, and 

also several profiles changed. Several profiles which were linear at 

0 M showed considerable curvature at 1.0 M and vice versa. This is 

due to the existence of alternate conformations (low energy excited 

states), that the residues can access. The residue wise details of these 

observations will be provided. 

Bi39 

The general form of fast oscillation model and its consequences 

for nitroxide EPR spectra interpretation in biological systems 

Tkachev, Yaroslav 

Engelhardt Institute of Molecular Biology, Molecular Biology, Moscow, 

Russian Federation 

Nitroxide spin probes are widely used for studying complex biological 

systems like proteins, lipid membranes, etc. Complex dynamics in these 

systems present at room temperature make unique interpretation of 

their EPR spectra a nontrivial task, as motions of different kind may 

have very similar effect on resulting spectrum. Simplifications can be 


made by splitting overall probe motion into several components based 

on their correlation times, and using some form of fast oscillation model 

[1] implying spin Hamiltonian averaging due to mechanical motion of 

the probe. 

We have developed a general form of this model allowing to classify 

these motions by their effect on EPR spectrum. The basis for our study 

was the two-motion model provided by V. Timofeev [1]. This model, 

being used in conjunction with our modified Temperature and Viscosity 

dependence (TVD) of EPR spectra technique [3], TSSE (Timescalebased 

State Splitting for Ensemble) model for timescale-based motion 

classification and describing ensemble inhomogeneity in fast motion 

model parameters, and, optionally, SLE (Stochastic Liouville Equation) 

for slow (in appropriate EPR frequency band) Brownian motion [2], gives 

a powerful method for studying structural and dynamical properties of 

spin probe neighborhood. This method has been successfully applied 

for studying the series of spin-labeled proteins (BSA, Fab fragment of 

human IgM) [3], as well as for studying of Barstar-Barnase complex 

formation [4] and lipid bilayers (with TEMPO used as probe). 

References 

[1] V. P. Timofeev, and B. A. Samarianov, J. Chem. Soc. PERKIN TRANS 

2. 1995, 2175-2181 

[2] D. J. Schneider, and J. H. Freed, Adv. Chem. Phys. 1989, 73, 387 

[3] V. P. Timofeev, Y. V. Tkachev, et al., J. Biomol. Struct. Dyn. 2005, 23, 

175-181 

[4] V. P Timofeev, V. V. Novikov, Y. V. Tkachev, et al. J Biomol Struct Dyn. 

2008, 25, 525-34. 

Bi40 

The sHSP αB-crystallin: NMR approaches for structural 

insights to its copper dependent chaperone-like activity and its 

interaction with the Alzheimers peptide Aβ 1-40 

Mainz, Andi; Jehle, Stefan; Oschkinat, Hartmut; Reif, Bernd 

Leibniz-Institut für molekulare Pharmakologie (FMP), Berlin, Germany 

Crystallins are the major structural proteins of vertebrate eye lenses 

maintaining lens transparency and proper refractive index. In contrast 

to αA-crystallin, localized primarily in the eye lens, αB-crystallin (αB) is 

distributed ubiquitously in several tissues and found to be elevated in 

brains of patients with Alzheimers (AD) and other neurological diseases. 

It assembles into polydisperse oligomers (~600 kDa), exhibiting a 

chaperone-like activity, which prevents unfolded proteins from either 

amorphous or amyloidogenic aggregation. However, the anti-fibrillar 

effect of αB against the β-amyloid peptide Aβ 1-40 can even increase 

the neurotoxicity of the latter. Interestingly, divalent metal ions like 

copper(II) have not only impact on AD plaque architecture, but also 

on the morphology and activity of αB. Here we report an approach, 

applying solid-state MAS NMR experiments on liquid samples of αB at 

high concentrations and low temperature in the presence of glycerol. 

Under the employed experimental conditions αB reveals extremely slow 

molecular tumbling in solution, thus enabling anisotropic interactions to 

be averaged out by MAS. The spectral quality is comparable to the one, 

reported for precipitated αB. Typical solid-state MAS NMR experiments 

can be recorded without precipitation, facilitating the investigation of 

protein ligand interactions. Complementary solution-state NMR data 

using truncated αB fragments is presented in order to verify the solidstate 

NMR results. 


Bi41 

Spectra of high dimensionality - towards easy signal 

assignment in proteins 

Zawadzka, Anna; Kazimierczuk, Krzysztof; Kozminski, Wiktor 

University of Warsaw, Chemistry Department, Warsaw, Poland 

Multidimensional experiments, invaluable in case of crowded spectra, 

may sometimes not ensure satisfactory results, when performed in a 

conventional way, where the evolution time space is sampled at equal 

intervals. The necessity of fulfilling the Nyquist Theorem in this case 

causes that having limited experimental time one has to compromise 

between resolution and spectral width. In such situation, when peaks 

overlap, it is hard to precisely evaluate chemical shifts. We have 

proposed the method which allows overcoming this problem. 

Random sampling of evolution time space, followed by Multidimensional 

Fourier Transform, provides spectra of very high resolution (it is possible 

to obtain natural linewidths) and enables performing experiments of 

high dimensionality (4D, 5D, 6D). The processing of such spectra takes 

advantage of resonance positions form spectra of lower dimensionality 

and thus results with a set of 2D spectra, which makes it very 

convenient to use. 

We propose several novel techniques which allow easy assignment of 

protein backbone signals. The experiments were performed on proteins 

of various sizes: from ubiquitin (76 residues) to Maltose Binding Protein 

(370 residues). 

Bi42 

Interaction between the amyloid β-peptide(1-40) and 

cyclodextrin dimers 

Wahlström, Anna 1 ; Danielsson, Jens 1 ; Jarvet, Jüri 2 ; Rebek, Julius Jr. 3 ; 

Gräslund, Astrid 1 

1 Stockholm University, Department of Biochemistry and Biophysics, 

Stockholm, Sweden; 2 The National Institute of Chemical Physics and 

Biophysics, Tallinn, Estonia; 3 The Scripps Research Institute, La Jolla, 

United States 

Alzheimer’s disease is a form of dementia affecting mostly elderly 

people. One hallmark of the disease is the presence of amyloid plaques 

in the brain, i.e. accumulations of dead and injured neurons and various 

macromolecules. The dominating component is the amyloid β-peptide 

(Aβ), a 39-42 peptide from the membrane spanning protein β-amyloid 

precursor protein. The peptide aggregates very easily and the proposed 

pathway involves formation of soluble oligomers which finally form 

insoluble fibrils. 

Cyclodextrin (CD) is a cyclic oligosaccharide with a varying number 

of glucopyranose units. It has a hydrophilic outer surface and a 

hydrophobic interior, properties that make it suitable for interaction 

with guest molecules with hydrophobic groups. The host property of 

CD is interesting in the context of Aβ aggregation, as a possible way to 

influence the aggregation process. 

The interaction between Aβ(1-40) and CD monomers (α-, β- and γ-CD 

with 6, 7 and 8 glucopyranose units, respectively) has been studied 

before, revealing interaction between only β-CD and the peptide. The 

interaction involves at least two binding sites and therefore we have 

continued the study with dimers of β-CD, connected by a linker of 

varying length and flexibility. The interaction between 50 µM unlabeled, 

15N fully labeled or 13C-15N fully labeled Aβ(1-40) and dimers in 

millimolar concentration was studied by NMR. The experiments included 

translational diffusion (PFG-LED) and 15N-1H and 13C-1H HSQC. 

Titration of increasing concentration of dimers to Aβ(1-40) demonstrated 

a decreasing diffusion coefficient, indicating a weak interaction between 

the peptide and the various dimers. The interaction was studied in 

detail by the HSQC experiments which revealed specific shifts of the 

resonances of the aromatic amino acid residues F4/Y10, F19 and F20. 

ABSTRACTS POSTER


Bi43 

Solution structure of the first two RNA recognition motifs of 

SiahBP 

Cukier, Cyprian D. 1 ; Diaz-Moreno, Irene 1 ; Hollingworth, David 1 ; Kelly, 

Geoff 2 ; Ramos, Andres 1 

1 MRC National Institute for Medical Research, Molecular Structure 

Division, London, United Kingdom; 2 MRC National Institute for Medical 

Research, MRC Biomolecular NMR Centre, London, United Kingdom 

SiahBP (Seven in absentia homologue protein-binding protein) is a 

multifunctional protein engaged in splicing and transcription regulation. 

The protein comprises three domains: the first two are canonical nucleic 

acids-interacting RNA recognition motifs (RRMs) while the third belongs 

to a protein-interacting RRM subfamily, called U2AF-homology motif 

(UHM domain). This study provides the solution structure of the first 

two RRM domains of SiahBP. The domains have a typical RRM topology 

(β1α1β2β3α2β4) where two alpha-helices are packed against a fourstrand 

anti-parallel beta-sheet. Further, two additional alpha-helical 

regions have been identified in the N-terminus of the construct and in 

the linker between the RRM domains. 

Bi44 

Lipid-protein nanodiscs: possible application in high-resolution 

NMR investigations of membrane proteins and membraneactive 

peptides 

Lyukmanova, Ekaterina; Shenkarev, Zakhar; Nekrasova, Oksana; 

Ovchinnikova, Tatiana; Arseniev, Alexander 

Shemyakin-Ovchinnikov Insitute of Bioorganic Chemistry, Moscow, 


Reconstituted nascent high-density lipoprotein particles or lipid-protein 

nanodiscs (LPNs) represent a fragment of lipid bilayer (~160 lipid 

molecules) stabilized by dimer of human apolipoprotein A1. These 

particles have discoid shape with characteristic dimensions 10x4 

nm. It is generally assumed that LPNs demonstrate better membrane 

mimicking properties than spherical detergent-containing micelles and 

small bicelles commonly used in high-resolution NMR spectroscopy 

experiments. The applicability of LPN based membrane mimetics 

in high-resolution NMR investigations of membrane proteins and 

membrane-active peptides embedded into LPN membrane was studied. 

The 15 N-labeled analogues of bacterial K+ channel KcsA (Streptomyces 

lividans) and antibiotic antiamoebin I (Aam-I, Emericellopsis minima) 

were incorporated into LPNs of various lipid compositions. The formation 

of stable complexes which undergo isotropic motion on the NMR time 

scale was confirmed by gel-filtration and 31 P-NMR spectroscopy. The 

2D 1 H- 15 N-correlation spectra were measured for KcsA in the complex 

with LPN containing DMPC and for Aam-I in LPNs based on DOPG, 

DLPC, DMPC, and POPC. The measured spectra were compared with 

those in detergent-containing micelles and small bicelles commonly 

used in high-resolution NMR spectroscopy of membrane proteins. 

The spectra recorded in LPN environment demonstrated similar signal 

dispersion but significantly increased line width. The observed 1 HN line 

width was about 50-80 Hz for membrane-associated fragments of the 

molecules and about 20-30 Hz for mobile solvent-exposed domains. 

The spectra of Aam-I embedded in LPNs containing phosphatidylcholine 

showed significant selective line broadening, thus suggesting exchange 

process(es) between several membrane-bound states of the peptide. 

15 N relaxation rates were measured to obtain the effective rotational 

correlation time of the Aam-I molecule. The obtained value (~40 nsec 

at 45°C) indicates the presence of additional peptide motions within 

the Aam-I/LPN complex. In order to give an example of structural 

information that can be gained in LPN environment the 3D 15 N-TROSY- 

NOESY spectrum of Aam-I/LPN complex was measured. 

50 

Bi45 

Functional insights into the catalytic mechanism of methionine 

sulfoxide reductase B1 through its structural analysis 

Aachmann, Finn L. 1 ; Sal, Lena s. 1 ; Kim, Hwa-Young 2 ; Gladyshev, Vadim 

N. 3 ; Dikiy, Alexander 1 

1 Norwegian University of Science and Technology, Department of 

Biotechnology, Trondheim, Norway; 2 Yeungnam University College of 

Medicine, Department of Biochemistry and Molecular Biology, Daegu, 

Republic of Korea; 3 University of Nebraska, Department of Biochemistry, 

Nebraska, United States 

Reactive oxygen species (ROS) can oxidize Met in proteins to a mixture 

of the R- and S-isomer of Met-sulfoxide (Met-SO). The Met-SO 

reductases (Msr) regenerate Met-SO back to Met in the presence of 

thioredoxin reductase (Trx). There are two different classes of Met-SO 

reductases: MsrA catalyses the reduction of the S-isomer while MsrBs 

reduce the R-form of Met-SO. 

Mammalian MsrB1 contains Sec in the active site while MsrB2/3 are 

cysteine homologs of MsrB1, having a Cys instead of Sec in their active 

site. It was proposed that Sec and Cys forms of MsrBs proteins use 

different mechanisms to catalyze Met-SO reduction. The main objective 

of this project is to distinguish different catalytic mechanisms of 

mammalian MsrBs proteins. 

Standard NMR experiments were used to obtain the resonance 

assignment on uniformly 13C/15N labeled reduced recombinant 

Sec95Cys MsrB1. The structure calculations were based on geometrical 

constraints derived from NOE and angles constraints obtained from 

TALOS. The MsrB1red structure is characterized as an overall â-fold 

protein consisting of eight antiparallel â-strands. The first 18 amino 

acids in the N-terminal form a highly mobile tail. 

The reaction mechanism for MsrB1 was studied by exposing it to both 

Met-SO (substrate) and Met-SO2 (inhibitor). The observed results 

indicate that the Cys4 at the protein N-terminal tail acts as the resolving 

Cys reacting with the oxidized catalytic Sec. The protein’s activity is then 

restored by reduction of the formed intramolecular seleno-sulpho bond 

by Trx. 

The pH dependence of the active site residues was investigated in order 

to establish whether the same level of structural perturbation is present 

in MsrB1 compared to other MsrBs. It was found that Cys95 has around 

2 pH unit lower pKa that a normal Cys. The accomplished studies 

together with the analysis of the data available for Cys-homologs of 

MsrB support the previously proposed suggestion that Sec-containing 

MsrB1 has different mechanism for Met-SO reduction in comparison 

with Cys-containing MsrBs. 

Bi46 

Segmental labelled AlgE4 - an alginate epimerase 

Buchinger, Edith 1 ; Aachmann, Finn L. 2 ; Skjåk-Bræk, Gudmund 2 ; Valla, 

Svein 2 ; Iwai, Hideo 3 ; Wimmer, Reinhard 1 

1 Aalborg University, Department of Biotechnology, Chemistry and 

Environ, Aalborg, Denmark; 2 Norwegian University of Science and 

Technology, NOBIPOL, Department of Biotehnology, Trondheim, Norway; 

3 University of Helsinki, Institute of Biotechnology, Helsinki, Finland 

Nuclear magnetic resonance (NMR) is a powerful method to determine 

protein dynamics and protein-ligand interactions. New NMR techniques 

have extended the size limit for the observation of NMR signals to 

over 100kDa, but signal overlap in large proteins can hinder spectral 

analysis. Isotopic labelling of only a segment of a large protein reduces 

the spectral complexity but still allows sequence-specific resonance 

assignment and functional investigations. 

One method of segmental isotopic labelling relies on the protein splicing 

activity of split inteins. Protein splicing needs no cofactors or reagents 

to excise an intervening sequence and ligate two flanking N-and 


C-terminal segments via a peptide bound. Additionally, the ligation of 

segments can be done in vivo or in vitro. 

In the bacterium Azotobacter vinelandii, a family of seven secreted and 

calcium-dependent mannuronan C-5 epimerases (AlgE1–7) has been 

identified. These epimerases are responsible for the epimerization of 

β-D-mannuronic (M) acid to α-L-guluronic acid (G) in alginate polymers. 

The epimerases consist of two types of structural modules, designated 

A [~350aa] (one or two copies) and R [~150aa] (one to seven copies). 

Each member of the AlgE-family produces a unique sequence of M 

and G subunits. The A-modules are catalytic active; the R-modules 

strongly enhance this activity although they don’t posses any catalytic 

activity. The smallest member of the family, AlgE4, consist of one A- and 

one R-module (A-R). The structure of the A-module of AlgE4 has been 

solved by X-ray crystallography and the structure of the R-module was 

solved by NMR spectroscopy. NMR studies on the R-module of AlgE4 

have shown that alginate also binds to the R-module. 

The function of the R-domains in the AlgE-family is to a large extent 

unknown. Our objective is to obtain an active segmentally labelled 

AlgE4 (14N-A-15N-R) for alginate binding studies to the R-module 

by NMR. For the protein splicing we used the naturally split intein of 

Nostoc punctiformae. Finally, a multiple segmentally labelled AlgE4 will 

be produced for interaction studies between both modules with and 

without alginate. 

Bi47 

NMR Study of Salmonella enterica serovar Typhimurium intact 

cells 

Zandomeneghi, Giorgia 1 ; Ilg, Karin 2 ; Aebi, Markus 2 ; Meier, Beat, H. 1 

1 ETH Zurich, Laboratory of Physical Chemistry, Zurich, Switzerland; 2 ETH 

Zurich, Institute of Microbiology, Zurich, Switzerland 

NMR spectroscopy can detect biomolecules directly in the cell, thus 

avoiding processes as isolation and purification, and providing a more 

realistic description than the one derived from in-vitro studies. The 

resulting spectra can be very complex, and the use of magic-angle 

spinning (MAS) is often required to obtain narrow lines, predominantly 

because MAS averages the magnetic-susceptibility variations present 

in such heterogeneous samples. Here we present a 1 H NMR study of 

the lipopolysaccharide (LPS) of the pathogenic bacterium S. enterica 

sv. Typhimurium, where we performed High-Resolution MAS NMR 

experiments directly on the intact cells. 

In Gram-negative pathogenic bacteria LPS, a major component of 

the bacterial outer membrane is an important virulence factor. In S. 

enterica, it consists of the O-antigen region, a polysaccharide chain 

constituted by up to 100 repeats of 4 to 6 monosaccharides, covalently 

linked to the membrane-anchored Lipid A core. The O-antigen is the 

most variable portion of LPS, varying between different strains and 

thus imparting antigenic specificity. LPS protects the bacteria from 

environmental stress and comprises one of the major pathogenassociated 

molecular pattern by which the immunological defence 

system of the host recognizes the invading bacteria as foreign. The 

characterization of LPS and its response to drugs, gene mutations 

and environmental effects can give insight into the mechanism 

of pathogenesis and assist to develop novel diagnostic tests and 

antimicrobial. 

We characterized LPS of S. enterica by NMR, being able to selectively 

observe the O-antigen. We determined the effects of growth phase, 

cell density and sample temperature on the spectral properties of LPS 

and the cellular metabolites. Two chemical shift variants for the LPS 

were detected in live cells, one corresponding to the signals of isolated 

LPS and one specific for the cellular microenvironment. Finally, we 

investigated the effect of mutations that affect LPS biogenesis, relating 

the spectral characteristics of various mutants to their pathogenicity and 

resistance to drugs. 


Bi48 

Folding core recognition and coarse-grained helix bundle 

model for the the 54.3 kDa sodium/proline symporter PutP of 

Escherichia coli 

Jeschke, Gunnar 1 ; Polyhach, Yevhen 1 ; Hilger, Daniel 2 ; Jung, Heinrich 2 

1 ETH Zürich, Lab. Phys. Chem., Zürich, Switzerland; 2 LMU München, 

Department Biologie I, München, Germany 

Despite recent advances, crystallization of membrane proteins is still 

tedious and often depends on serendipity. Although structures of several 

related proteins could be solved during the past few years, among them 

the sodium/galactose symporter vSGLT of Vibrio parahaemolyticus from 

the same family, attempts to crystallize the sodium/proline symporter 

PutP of E. coli have failed so far. All the presumably related transporters 

share the fold of a core of ten transmembrane helices, while between 

2 and 4 auxiliary transmembrane helices are located in different ways 

with respect to this core. In previous work we had established that one 

of the core helices is significantly kinked, having the same shape as the 

presumably equivalent helix in the leucine transporter LeuT Aa of Aquifex 

aeolicus [1]. 

Based on double electron electron resonance (DEER) distance 

measurements between spin labels attached to helix ends, we have 

attempted to model the helical bundle of PutP. Using a labeling pattern 

based on 13 cytoplasmic and 13 periplasmic sites, we have gathered 

19 cytoplasmic, 23 periplasmic, and 4 transmembrane distance 

restraints. Due to uncertainties in label orientations and conformations 

these restraints are not sufficient to derive a well defined model of 

the helix bundle from scratch using distance geometry. Such a model 

can be obtained after adding template restraints derived from the tenhelix 

core of the vSGLT crystal structure. Our approach distinguishes 

with high certainty between structurally homologous and structurally 

unrelated core templates. Hence, the fold of the core can be reliably 

recognized. Furthermore, arrangement of the 3 auxiliary helices of 

PutP with respect to the core is established. Prerequisites for extending 

this approach to structure determination from scratch, without a core 

template, are discussed. 

[1] D. Hilger, Y. Polyhach, H. Jung, G. Jeschke, Biophys. J., 96, 217-225 

(2009). 

Bi49 

Prokaryotic Reggie-associated NfeD Proteins adopt the OB-fold 

and localize into lipid microdomains 

Möller, Heiko M. 1 ; Walker, Christina A. 1 ; Witte, David 1 ; Hinderhofer, 

Markus 2 ; Boos, Winfried 2 ; Dempwolff, Felix 3 ; Graumann, Peter 3 

1 University of Konstanz, Chemistry, Konstanz, Germany; 2 University of 

Konstanz, Biology, Konstanz, Germany; 3 University of Freiburg, Biology, 

Freiburg, Germany 

Reggie/Flottilin proteins are upregulated during neuronal regeneration 

and play crucial roles in various cell signaling processes. We have 

analyzed the genomes of prokaryotic organisms and found a large 

number of reggie-like genes. Remarkably, these reggie-like genes are 

very often accompanied by a gene of another transmembrane protein 

belonging to the diverse NfeD family, both of them organized in the 

same operon structure and thus co-expressed and co-regulated. This 

is a clear indication for a functional relationship of both genes and 

potentially even for a physical interaction of their respective proteins. 

In B. subtilis, the reggie-like operon is regulated by the SigmaW-factor 

leading to enhanced expression under conditions of cellular stress, like 

osmotic shock or presence of cell envelope-compromising antibiotics. 

It appears that reggie-like proteins (RLPs) and their associated partner 

proteins (RAPs) altogether act in response to these stress conditions. 

As a first step on the way to elucidate the functional network of these 

ABSTRACTS POSTER


proteins, we have determined 3D structures in solution of the cytosolic 

portion of three different RAPs. Despite very low homology at the level 

of primary structure, all RAPs share a structural core composed of a 

five-stranded beta-barrel, well known from the diverse class of OB-fold 

proteins that are usually involved in oligonucleotide and oligosaccharide 

binding. However, considering the pattern of conserved amino acids 

RAPs appear to be more likely involved in protein-protein interactions. 

These interactions may be responsible for co-localizing RAPs and RLPs 

in lipid microdomains as determined by fluorescence microscopy. 

Bi50 

Solution structure of IPSE/alpha-1 

Meyer, Helge 1 ; Tripsianes, Konstantinos 1 ; Madl, Tobias 1 ; Blindow, Silke 2 ; 

Bade, Steffen 3 ; Barths, Daniela 2 ; Schramm, Gabriele 2 ; Sattler, Michael 1 

1 Helmholtz Zentrum Munich, Biomolecular NMR, Munich, Germany; 

2 Research Centre Borstel, Cellular Allergology, Borstel, Germany; 

3 Research Centre Borstel, Mucosaimmunology, Borstel, Germany 

Shistosomiasis, which amongst others is caused by the parasite 

Schistosoma mansoni, has been recognized as the most important 

worm infection in terms of morbidity and mortality. During infection with 

this parasite the deposition of schistosome eggs subsequently leads 

to a T helper type 2 (Th2) cell response triggered by the production of 

Interleukin 4. The interleukin-4-inducing principle from Schistosoma 

mansoni eggs (IPSE/alpha-1) which is secreted from Shistosoma 

mansoni eggs, has been identified as the major antigenic compound, 

although the mechanism of induction of immune response remains 

unclear. 

Here we report the 3D structure of IPSE/alpha-1, which we have 

determined by using solution NMR spectroscopy. The protein structure 

classifies IPSE as a non-lens member of the gamma-crystallin 

superfamily, albeit the very low sequence homology with the known 

crystallin members. Nevertheless , the structure is highly conserved 

consisting of two Greek-key motifs with a basic 2-beta-sheet 

overall fold. What distinguishes IPSE from the other crystallins is the 

extended variable loop that links the two Greek-key motifs and the fold 

dependence on the formation of three disulfide bridges identified from 

the carbon chemical shifts of the CYS residues and other biophysical 

methods. Current studies aim at elucidating the functional details of Th2 

activation using the available structural information. 

Bi51 

Production and NMR structural studies of the second 

transmembrane domain from human wild-type and mutant 

melanocortin-t receptor 

Yu, Jeong-A 1 ; Gang, Ga-Ae 2 ; Park, Tae-Joon 2 ; Kim, Yongae 2 

1 Chosun University, Department of Science Education (Chemistry Major), 

Gwangju, Republic of Korea; 2 Hankuk University of Foreign Studies, 

Department of Chemistry, Yong-In, Republic of Korea 

It has been suggested that normal melanocortin 4 receptor among five 

subtypes (MC1R-MC5R) increases energy expenditure and decreases 

food intake, and genetic disruption of MC4R causes severe early onset 

obesity. Therefore, MC4 receptors may be ideal pharmacological 

targets for treating disorders such as obesity and anorexia. MC4R is 

membrane-bound protein that transverse the lipid bilayers of the cell 

membrane, so it is difficult to express and characterize the membranebound 

three-dimensional structure by using conventional solution NMR 

and X-ray crystallography. 

In this study, we expressed and purified the wild-type TM2 and mutant 

TM2 peptides of MC4R. We successfully cloned and optimized the 

expression condition and purified the wild-type TM2 and mutant TM2 

peptide of MC4R. Approximately, the yield was about 200mg/1L growth 

52 

with a fusion partner. The two recombinant peptides were characterized 

by tris-tricine PAGE, mass spectroscopy, and HSQC nmr spectroscopy. 

In addition, their initial structural data was obtained and compared with 

solution NMR spectroscopy and solid-state NMR spectroscopy in the 

membrane-like environments. 

Reference 

1. Protein Expression and Purification 62 (2008) 139 

2. Nature 385 (1997) 165 

3. Diabetes 52 (2003) 2984 

Bi52 

Folding and topology of ribosomal protein S6 and permutant 

investigated with 2D NMR experiments 1 H N - 2 H N Exchange 

Lind, Jesper; Haglund, Ellinor; Oliveberg, Mikael; Mäler, Lena 

Stockholm University, Biochemistry and Biophysics, Stockholm, Sweden 

The ribosomal protein S6 is a two state model á/â protein exceptionally 

well suited for folding studies. Previously five topological variants of S6 

were constructed to explore how entropy changes influence the folding 

process and folding cooperativity. In these variants the mutual orders 

of secondary structure elements have been altered without changing/ 

effecting the final native structure of the protein. Only the loops are 

connected in new ways by circular permutation [1,2]. 

In this work the wild-type S6 (S6wt) and the P54-55 permutant have 

been investigated with high resolution NMR spectroscopy. 1 H N - 2 H N 

exchange rates were monitored over 60 hours by recording HSQC of 

lyophilized S6wt and permutant P54-55 dissolved in 2 H 2 O. In addition 

SOFAST-HMQC was used to study H N exchange on a faster time-scale. 

2D experiments with varying presaturation delays were performed on 

the proteins in H 2 O solution in order to capture the immediate amide 

proton exchange. 

This poster will present results of the 1 H N - 2 H N exchange experiments 

of the ribsomal protein S6wt and the P54-55 permutant together with 

the conclusions that can be drawn about the possibilities to study the 

folding mechanisms of these small α/β proteins. It has previously been 

shown that the wild type protein and the permutant fold differently 

involving two distinct foldons [3]. The present work shows that both 

S6wt and the P54-55 permutant displays increased exchange rates 

in the same part of the structure indicating that there is an inherent 

flexibility in S6, which is not correlated to the folding process. 

1 Lindberg, M et al. (2001) J Mol Biol, 314, 897-900 

2 Lindberg, M. et al. (2002) Nat Struct Biol, 9, 818-22. 

3 Haglund, E. et al. (2008) J Biol. Chem, 283, 27904-27915 

Bi53 

Zinc and copper binding to Amyloid-β peptide: pH dependence 

of binding strength and specificity of binding site 

Ghalebani, Leila; Wahlström, Anna; Gräslund, Astrid 

Stockholm University, Department of Biochemistry & Biophysics, 

Stockholm, Sweden 

It has been shown that aggregation of the amyloid-β peptide (Aβ, 40- 

42 residues) is involved in the pathogenesis of Alzheimer’s disease 

(AD). Furthermore, there is evidence that binding of divalent metal 

ions such as zinc and copper to the peptide affects the aggregation 

in a non-trivial manner, either inducing or preventing the peptide 

aggregation depending on metal concentration. Here we have studied 

the interactions of zinc and copper with Aβ (1-40) at low concentration 

of the peptide, using fluorescence and NMR spectroscopy as the 

main methods. The detailed molecular effects of low concentrations 

of zinc and copper binding on the full length Aβ (1-40); is explored 


under various acidic conditions. In agreement with previous studies, 

we observe competitive binding of Zn 2+ and Cu 2+ at physiological pH; 

both ions share a specific major binding site in the N-terminal part 

of the peptide and have similar binding affinities (KD in the range of 

micromolar). At lower pH (pH 5.5), however, our NMR results indicate 

that there is no specific binding site for zinc, whereas the specific 

binding site for copper remains the same. The results are compared 

with the effects of the two metal ions on the Aβ aggregation process. 

Bi54 

Structural analysis of EPS from Pedioccocus parvulus by NMR 

and molecular modeling 

Calle, Luis Pablo 1 ; Jiménez-Barbero, Jesús 1 ; Cañada, Francisco Javier 1 ; 

López, Paloma 2 ; Werning, Laura 2 

1 CIB-CSIC, Chemical and Physical Biology, Madrid, Spain; 2 CIB-CSIC, 

Molecular Microbiology and Infection Biology, Madrid, Spain 

A number of exopolysaccharides β-glucan type play an important role 

in the rheology and texture of fermented foods and have also shown 

prebiotic properties. In addition, the (1-3)-β-D-glucans can promote 

antitumor and antimicrobial activity, by activating macrophages, other 

white blood cells or dendritic cells. 

It has been shown that the lactic acid bacteria Pedioccocus parvulus 

2.6 synthesizes an extracellular exopolisaccharide with thickening 

properties, that has been characterized as a linear β (1-3) homopolymer 

of D-glucose substituted every two glucose residues by another 

β-glucose unit on the 2 position. 

The NMR characterization and assignment1 of this β-glucan and its 

rheological properties have been previously analysed. 

This exopolisaccharide has been produced and secreted (300 mg 

L-1) by an uncapsulated recombinant Lactococcus lactis strain. This 

bacteria expresses the P. parvulus glycosyltranferase responsible for the 

β-glucan biosynthesis. 

The size of this polymer and the repeated trisaccharidic moiety make 

difficult the study based on the analysis of NOE spectra. Our current 

strategy tries to elucidate the structure and the conformation of this 

exopolysaccharide in water thanks to RDC data and computational 

evaluation. 

Bi55 

Modelling of time series Microarray data using dynamic 

Bayesian network 

Manoj, Manoj; KGS, K.G.Srinivasa; Seema, S. 

MSRIT, Bangalore, CSE, Bangalore, India 

Abstract-Gene Regulatory Network represents how the genes interact 

with each other. Using genetic network modelling, it is possible to 

explain the cell functions at molecular level. DNA microarrays can 

measure the expression levels of thousands of genes simultaneously. 

Two steps method adapted to model large-scale Gene Regulatory 

Networks using time series microarray data. Firstly, genes are clustered 

based on existing biological knowledge (Gene Ontology annotations) 

and then a dynamic Bayesian network applied in order to model causal 

relationships between genes in each cluster. Finally the learned subnetworks 

are integrated to make a global network. This project aims at 

inferring the regulatory network that provides us the interaction between 

the various genes. Our aim is to apply data mining technique to gene 

expression data and infer regulatory network for various experiments, 

which include experiments in good and bad conditions using the 

information available in Gene Ontology 


Bi56 

Coffea arabica arabinogalactan-protein from instant coffee 

Matulova, Maria 1 ; Capek, Peter 1 ; Navarini, Luciano 2 ; Suggi-Liverani, 

Furio 2 

1 Institut of Chemistry, Center for Glycomics, SAS, Phys. Analytical Dept., 

Bratislava, Slovakia; 2 Illycaffè s.p.a., Research & Innovation, Trieste, Italy 

Carbohydrate components in coffee beans arabinogalactan-protein 

(AGP) represent about 17% of dry weight. They are very important as 

the roasted coffee aroma precursor and they play an important role in 

the determination of some coffee brews functional properties. 

Carbohydrate part of the AGP is primarily O-linked to Hyp / (Ser/Ala) 

as type II arabino-3,6-galactans. According the proposed structure, 

the backbone composed of 1,3-linked betaGal is highly substituted at 

O6 by 1,6-linked betaGal side chains of different length branched at 

O3 by alphaAraf, Gal, Rha and GlcA. It has a highly branched structure 

[1]. Up to now little attention has been dedicated to the study of AGPs 

modifications during an instant coffee preparation from roasted Coffea 

arabica beans [2,3]. 

Present study reports on the isolation and structural features of AGP 

from instant coffee powder prepared from roasted Coffea arabica 

beans. Results of chemical and spectroscopic methods suggest a high 

degree of AGP destruction after a drastic industrial processing when 

compared with AGP from green coffee. Its molecular size was markedly 

reduced to about 5 400. During this process linkages in the backbone 

and side chains were broken. The quantity of non-substituted 1,3-linked 

betaGal units of the backbone increased in comparison to 1,3,6-linked 

ones. Araf residues were found to be partly hydrolyzed on internal 

1,6-linked Gal residues of side chains and were missing on their 

terminal ends. No GlcA and rhamnose was detected. In addition, the 

intensity of signals in NMR spectra due to terminal 1,3-linked betaGal of 

the backbone as well as due to alpha,betaGal units of the reducing ends 

indicates the presence of polysaccharides of lower molecular weights in 

the mixture. 

Aknowledgments: Project has been supported by the IllyCaffe grant, 

VEGA Grant No. 2/0155/08, the Slovak State Program Project No. 

2003SP200280203. 

References 

[1] Regwell R. et al.; Foods Food Ingredients J. Jpn., 2006, 221 (1), 

38-46. 

[2] Wolfrom, L.; Anderson, E.; J. Agr. Food Chem., 1967, 15, 687-689. 

[3] Capek, P., Matulova, M.; Navarini, L.; Suggi-Liverani F.; J. Food and 

Nutr. Res., 2009, in press. 

Bi57 

Distinct dimeric conformations of EphA transmembrane 

domains reveal diversity of transmembrane helix packing 

among receptor tyrosine kinases 

Mayzel, Maxim 1 ; Bocharov, Eduard 1 ; Mineev, Konstantin 1 ; Goncharuk, 

Marina 1 ; Volynsky, Pavel 2 ; Efremov, Roman 2 ; Arseniev, Alexander 1 

1 Institute of Bioorganic Chemistry, Laboratoy of biomolecular NMR 

spectroscopy, Moscow, Russian Federation; 2 Institute of Bioorganic 

Chemistry, Laboratoy of biomolecular modeling, Moscow, Russian 

Federation 

The Eph receptors, largest family of receptor tyrosine kinases (RTK), 

and their membrane-bound ephrin ligands control a diverse array of 

cell-cell interaction in the developing and adult organisms. During signal 

transduction, the Eph receptors are involved in lateral dimerization 

and subsequent oligomerization processes within plasma membrane 

microdomains. While accumulating evidences reveal that the proper 

lateral dimerization of transmembrane (TM) domains of RTK is required 

for biochemical signal transduction across plasma membrane, the 

ABSTRACTS POSTER


functional role of the TM domains in the Eph receptor association was 

so far unclear. 

The structural-dynamic properties of the homodimeric TM domains of 

EphA1 and EphA2 receptors were investigated with the aid of solution 

NMR in lipid bicelles and molecular dynamics energy relaxation in 

explicit lipid bilayer. The distinct right- and left-handed parallel helixhelix 

packing were revealed for the TM domains embedded into the 

DMPC/DHPC bicelles. The EphA1 TM domain self-associates in a righthanded 

parallel α-helical bundle (544-569) through the N-terminal 

glycine zipper motif. In turn, the left-handed parallel EphA2 TM 

helices (535-559) interact through the extended heptad repeat motif, 

whereas the additional glycine zipper motif is not employed, enabling 

the receptor to adopt more than one dimeric conformation in plasma 

membrane. 

These findings serve as an instructive example of the diversity of 

TM domain formation within the same family of protein kinases and 

appear to favor the assumption of so-called rotation-coupled activation 

mechanism of Eph receptor signaling. In addition, our data suggest 

that the TM domains of Eph receptors and surrounding lipid bilayer do 

not merely play a passive role in signal transduction, but can provide 

an additional driving force for underlying conformational transition and 

extra specificity for the Eph receptor dimerization and clustering. 

The structural investigation of the EphA TM domain association helps 

to understand the underlying mechanisms of the Eph-ephrin signal 

transduction and provides a basis to control the receptor kinase activity, 

especially in pathological states of organism. 

Bi58 

Structural Venomics: An integrated platform for highthroughput 

NMR structure determination of toxins 

Mobli, Mehdi; Low, Check-Fong; Saez, Natalie; King, Glenn F. 

University of Queensland, Institute for Molecular Bioscience, Brisbane, 

Australia 

Animal toxins have been refined over millions of years for stability, 

activity and specificity, and it is becoming increasingly clear that they 

are ideal substrates for the modulation of proteins involved in important 

biological processes. However, to further our understanding of this 

important class of molecules a platform is required for their high-yield 

production, purification and structural characterization. The latter is of 

particular importance in toxins as their tertiary structure is conserved 

much more strongly than their amino acid sequence. 

Although NMR is an ideal tool for structural characterization of these 

relatively small proteins, the procedures used for determining protein 

structures using NMR are time consuming and labor intensive, which 

substantially reduces throughput. Thus, we have developed a robust 

platform for the high-throughput structure determination of isotopically 

labelled recombinant peptide toxins. The labelled proteins are produced 

recombinantly in the periplasm of E. coli where access to the disulfidebond 

processing machinery allows for high yield of correctly folded 

product. The NMR structure of the labelled material is then determined 

using a pipline we have developed, called “Automated Structural 

Analysis of Proteins by NMR” (ASAP-NMR), which allows high quality 

structural characterization of entire venomes at an unprecedented rate. 

The core of this platform is based on automated maximum entropy 

(MaxEnt) reconstruction of non-uniformly sampled NMR data 1 which 

reduces the NMR data-acquisition time from over a month to under a 

week. Data analysis is subsequently performed in a highly automated 

manner using probabilistic models 2 , and the structural restraints are 

translated into high-resolution NMR structures using the automated 

structure calculation program CYANA 3 . 

1. Mobli, M., Maciejewski, M. W., Gryk, M. R., and Hoch, J. C. (2007) 

Nat Methods 4, 467-468 

2. Bahrami A., Assadi A. H., Markley J. L. and Eghbalnia H. R. (2009) 

54 

PLoS Comput. Biol. 5(3): e1000307 

3. Guntert, P. (2004) Methods Mol Biol 278, 353-378 

Bi59 

The N-terminal domain of a RECQ-type helicase resembles a 

DNA binding motif 

Ohlenschlaeger, Oliver 1 ; Schneider, Annerose 2 ; Grosse, Frank 2 ; 

Pospiech, Helmut 2 ; Goerlach, Matthias 1 

1 Leibniz Institute for Age Research, Biomolecular NMR Spectroscopy, 

Jena, Germany; 2 Leibniz Institute for Age Research, Biochemistry, Jena, 

Germany 

The RecQL4 helicase, a member of the RecQ family of DNA helicases, is 

involved in the maintenance of genome integrity and in DNA replication. 

Mutations in the human RecQL4 gene cause the Rothmund-Thomson, 

RAPADILINO and Baller-Gerold syndromes. These diseases are 

characterised by increased cancer rates and symptoms of premature 

aging such as early development of cataracts and loss of hair. Mouse 

models and experiments have proven the N-terminal domain of RecQL4 

to be vital for cell growth. We have expressed the N-terminal domain of 

RecQL4 in E. coli (RecQL4N) and established a high-yield purification 

scheme to allow structure determination by heteronuclear liquid-state 

NMR spectroscopy. The first high-resolution structure of the human 

RecQL4 N-terminal domain carries an overall helical fold similar to 

a class of DNA-binding proteins albeit lacking significant sequence 

homology. 

Bi60 

Redox-dependent domain rearrangement of protein disulfide 

isomerase coupled with exposure of its substrate-binding 

hydrophobic surface 

Maeno, Aya 1 ; Nakano, Michiko 2 ; Kamiya, Yukiko 2 ; Serve, Olivier 2 ; 

Sasakawa, Hiroaki 2 ; Inaba, Kenji 3 ; Kurimoto, Eiji 4 ; Kato, Koichi 2 ; Kajino, 

Tsutomu 5 ; Nakasako, Masayoshi 6 

1 Nagoya City University, Nagoya, Japan; 2 Institute of Molecular Sciences, 

okazaki, Japan; 3 Kyushu University, Fukuoka, Japan; 4 Nagoya City 

University, Graduate School of Pharmaceutical Sciences, Nagoya, Japan; 

5 Toyota Central R&D, Nagakute, Japan; 6 RIKEN Harima Institute, Sayo- 

gun, Japan 

Protein disulfide isomerase (PDI) is a major protein in the endoplasmic 

reticulum, operating as an essential folding catalyst and molecular 

chaperone for disulfide-containing proteins, by catalyzing the formation, 

rearrangement, and breakage of their disulfide bridges. This enzyme 

has a modular structure with four thioredoxin-like domains, a, b, b’ 

and a’ along with a C-terminal extension. The homologous a and a’ 

domains contain one cysteine pair in their active site directly involved 

in thiol-disulfide exchange reactions, while the b’ domain putatively 

provides a primary binding site for unstructured regions of the substrate 

polypeptides. Here we report a redox-dependent intramolecular 

rearrangement of the b’ and a’ domains of PDI elucidated by combined 

use of NMR and small-angle X-ray scattering (SAXS) methods. 

We first determined the solution structures of the b’ and a’ domains 

from Humicola insolens, a thermophilic fungus, and further proposed 

a three-dimensional structure model of these domains linked to each 

other by using the SAXS and NMR data (NOE and RDC experiments run 

at 920 MHz) as conformational restraints for the Xplor-NIH package. Our 

NMR data (1H-15N HSQC titrations) also showed that the substrates 

bound to a hydrophobic surface spanning these two domains, which 

became more exposed to the solvent upon oxidation of the active 

site of the a’ domain as shown by the increased H/D exchange rates. 

Fluorescence experiments with a hydrophobic probe further confirmed 

the increased exposure to solvent. Moreover, the SAXS profiles revealed 


that oxidation of the a’ active site causes segregation of the two 

domains. Based on these data, we propose a mechanistic model of PDI 

action; the a’ domain transfers its own disulfide bond into the unfolded 

protein accommodated on the hydrophobic surface of the substratebinding 

region, which consequently changes into a “closed” form 

releasing the oxidized substrate. 

Bi61 

Protein domain organization studies using residual dipolar 

couplings: Filamin A immunoglobulin-like domains 

Koskela, Outi 1 ; Permi, Perttu 2 ; Jiang, Pengju 3 ; Campbell, Iain 3 ; Ylänne, 

Jari 4 ; Kilpeläinen, Ilkka 1 

1 University of Helsinki, Department of Chemistry, Helsinki, Finland; 

2 University of Helsinki, Institute of Biotechnology, Helsinki, Finland; 

3 University of Oxford, Department of Biochemistry, Oxford, United 

Kingdom; 4 University of Jyväskylä, Department of Environmental and 

Biological Science, Jyväskylä, Finland 

Many protein sequences are composed of multiple structural units 

i.e. protein domains or modules. Even though separated domains are 

usually able to fold independently they are not necessarily structurally 

and functionally isolated. Protein modules may assemble to form more 

complex structures and they can function cooperatively. Most of the 

protein structures solved so far are isolated protein domains. In order 

to fully understand function of a multi-domain protein it is necessary to 

know how the domains are organized and how they interact. Residual 

dipolar couplings (RDCs) provide long-range structural information for 

NMR spectroscopic structure determination of biomolecules, even for 

larger structures for which the short-range NOE restraints may fail. 

Orientational nature of RDC constraints makes them especially useful in 

protein domain organization studies. 

We have used the RDC approach in structural characterization of filamin 

A immunoglobulin-like domains (IgFLNa). Filamin A is an actin binding 

and cross-linking protein which also interacts with a vast array of other 

proteins. Filamin A contains an N-terminal actin-binding domain and 

24 IgFLNs. Our goal is to find out the domain packing of the IgFLNa 

domains 16-21, which are involved in many of the filamin interactions. 

High molecular weight (60 kDa) of the protein necessitates use of 

perdeuteration and TROSY-techniques to suppress the unfavourable 

relaxation effects. We have previously solved the structures of 20 

kDa IgFLNa16-17 (PDB:2K7P) and 18-19 (PDB:2K7Q) using NMR 

spectroscopy and the crystal structure of IgFLNa19-21 (PDB:2J3S) 

is also available. Using the known module structures and RDC-based 

orientational constraints we have been able to elucidate the mutual 

organization of the six modules of IgFLNa16-21. 

Bi62 

Comparison of MD derived and experimental order parameters 

on M-PMV matrix protein 

Srb, Pavel 1 ; Lang, Jan 1 ; Vlach, Jiri 2 ; Prchal, Jan 3 ; Ruml, Tomas 3 ; Hrabal, 

Richard 2 

1 Charles University in Prague, Math and Physics Faculty, Prague, 

Czech Republic; 2 Institute of Chemical Technology, Laboratory of 

NMR spectroscopy, Prague, Czech Republic; 3 Institute of Chemical 

Technology, Department of Biochemistry and Microbiology, Prague, 

Czech Republic 

Mason-Pfizer monkey virus (M-PMV) belongs to the genus of 

betaretroviruses, in which the matrix protein (MA) plays the essential 

role in certain stages of their life cycle (e.g. in assembly, transport and 

budding of new viral particles). Several single or double point mutants 

are known to cause dramatic changes in the virus life cycle [1]. In 

particular, the single point mutation R55F in MA redirects the assembly 

of viral capsid towards the plasma membrane rather than to cytoplasm, 

which is the place of assembly of the wild type form (WT). 

The NMR assignment and three-dimensional structures of R55F and WT 

were solved by our group [1,2,3]. Both structures are composed of four 

α-helices, however, the respective orientation of helices 1, 2 and 3, 4 

is nearly opposite. This causes different accessibility of CTRS sequence 

for interaction with a molecular motor. Therefore, we attempt a detailed 

motional analysis of the loop connecting helices 2 and 3. 

The standard Lipari-Szabó analysis of 15 N relaxation data is 

complemented with analysis of 120 ns long MD trajectories. Several 

methods of processing of the MD data for N-H vectors motion can 

be found in literature. Here we namely choose those proposed by 

Brüschweiler [4] and Zidek [5]. These methods will be compared and 

discussed. 

References 

1 J. Vlach, J. Lipov, V.Veverka, M.Rumlova, T. Ruml, R.Hrabal, J.Biomol. 

NMR, 2005, 4, 381-2 

2 J. Vlach, et al, ,PNAS, 2008, 105, 30, 10565-10570 

3 J. Vlach, P. Srb, J. Prchal, M. Grocky, J. Lang, T.Ruml and R. Hrabal, 

J.Mol.Biol., 2009, in press 

4 J. Prompers, R. Brüschweiler, ,J.Am.Chem.Soc., 2002, 124, 4522- 

4534 

5 P. Macek, et. al., J.Phys.Chem.B, 2007, 111, 5731-5739 


Bi63 

Probing salt-bridge induced stability of TC5b variants 

Rovó, Petra 1 ; Farkas, Viktor 1 ; Hegyi, Orsolya 2 ; Csikós, Orsolya 2 ; Gábor, 

Tóth 2 ; Perczel, András 1 

1 Eötvös Loránd University, Laboratory of Structural Chemistry and 

Biology, Budapest, Hungary; 2 University of Szeged, Department of 

Medical Chemistry, Szeged, Hungary 

The Trp-cage miniprotein TC5b (NLYIQ WLKDG GPSSG RPPPS) is a 

20 residue long protein notable for its protein-like 3D fold in quasi 

physiological conditions [1]. In its structure an -helix, a 3 10 -helix and 

a polyproline II helix shield the central Trp6 residue. The structure is 

stabilized through a series of hydrophobic interactions organized around 

the Trp6 and through the salt-bridge formed between the residue Asp9 

and Arg16. 

Herein, we have investigated the molecular properties and stabilization 

efficacy of the Asp9-Arg16 salt-bridge by NMR and ECD techniques. 

The starting model was the previously designed and salt-bridgeoptimized 

variant TC6b (TC5b_D9E) in which a methylene-group was 

“inserted” into the Asp9 side-chain [2]. In this study we systematically 

changed the position of the previously inserted methylene-group and 

examined its effect on the globular structure stability. To probe the saltbridge 

induced stability, solvent- and heat-induced denaturation studies 

were conducted on the TC5b salt-bridge variants and deletion mutants. 

Our data deliver a rather comprehensive stabilization power of the 

examined salt-bridge. 

Our results strongly suggest that a salt-bridge in any protein should be 

evaluated in context with the surrounding residues as the interaction 

between the charged groups is at the same magnitude as the 

background interaction of the side-chains. As in case of the examined 

mutants an optimal H-bonding network could enhance the effectiveness 

of a Coulombic interaction between oppositely charged residues. 

[1] Neidigh, J. W. Fesinmeyer, R. M., Andresen, N. H., Nat. Struct. Biol. 

2002 9, 425-430. 

[2] Hudáky, P., Stráner, P., Farkas, V., Váradi, G., Tóth, G., Perczel, A., 

Biochemistry, 2008, 47, 1007-1016. 

ABSTRACTS POSTER


Bi64 

Interactions of the nucleotide binding domains of the human 

ABC transporter ABCC1 during the catalytic cycle 

Marchand, Philipp; Jacquet, Eric; Sizun, Christina 

ICSN-CNRS, Gif-sur-Yvette, France 

ABC transporters constitute a large family of membrane proteins, that 

are involved in various cellular functions. Their human member MRP1 

/ ABCC1 is associated to resistance of cancer cells to chemotherapy 

by extruding a large variety of molecules. Like all members of the ABC 

family, ABCC1 contains two nucleotide binding domains (NBD) and 

two transmembrane domains (TMD). While the TMDs are responsible 

for substrate translocation, the NBDs are energizing transport by ATP 

hydrolysis. The transport mechanism requires specific interactions 

between all four domains. 

Previous work has shown that the isolated NBDs of MRP1 / ABCC1 

show rather low hydrolytic activity and lack cooperativity, but undergo 

transitory interactions, as shown by NMR. These interactions show 

differences depending on different states of the catalytic cycle (Ramaen 

et al. 2005). In parallel the three-dimensional structure of NBD1 in 

complex with MgATP was obtained by X-ray diffraction measurements 

(Ramaen et al. 2006). 

Further characterization suggests that additional interaction partners 

are required for the stable formation of the heterodimer NBD1 – NBD2. 

Based on the three-dimensional structure of the full-length bacterial 

transporter Sav1866, cytoplasmic loops between transmembrane 

helices of the TMDs act as potential interaction partners, which remains 

to be characterized for ABCC1. 

However, so far it has not been possible to obtain NBD2 samples 

suitable for NMR experiments, in contrast to NBD1. New attempts 

of protein production and purification have been made to solve this 

problem and to enable us to investigate the interaction of NBD2 

with mimicking peptides of helical loop regions and finally the 

characterization of the NBD1 – NBD2 heterodimer in the presence of 

nucleotides by NMR. 

Ramaen et al. (2005) “Attempts to characterize the NBD heterodimer 

of MRP1: transient complex formation involves Gly771 of the ABC 

signature sequence but does not enhance the intrinsic ATPase activity.” 

Biochem J 391(3): 481 

Ramaen et al. (2006) “Structure of the Human Multidrug Resistance 

Protein 1 Nucleotide Binding Domain 1 bound to Mg2+/ATP Reveals a 

Non-productive Catalytic Site.” J Mol Biol 359(4): 940 

Bi65 

Solution NMR spectroscopy studies of unfolded Cu/Zn- 

Superoxide dismutase 

Szpryngiel, Scarlett; Oliveberg, Mikael; Mäler, Lena 

Stockholm University, Dept. of biochemistry and biophysics, Stockholm, 

Sweden 

Human Cu/Zn Superoxide dismutase (SOD1) is an enzyme that 

acts as a radical scavenger, protecting the cell from toxic oxygen 

species. SOD1 is a homodimeric protein containing 153 amino acid 

residues. The protein has one copper (catalytic role) and one zinc ion 

(structural role) bound per subunit. At least 111 missense mutations 

in the gene encoding SOD1 are associated with a familial form of the 

neurodegenerative disease ALS (fALS). It is, however, unclear how these 

protein variants are involved in the disease mechanism since they 

are not affecting the dismutase activity, i.e. it is not a loss-of-function 

mechanism. The disease is instead thought to be due to a gain of 

toxic function coupled to some misfolding event and fALS is therefore 

classified as a misfolding disease. Cytoplasmic SOD1 aggregates are 

commonly found in fALS patients but the cytotoxicity of SOD1 seems 

56 

to be due to species arising earlier in the folding pathway and not the 

aggregates themselves. These species are monomeric without any 

metals bound (apo-SOD1). It is unknown at which stage of the folding 

process the metallation takes place. 

The aim of this project is to study early folding events of SOD1 and 

capture species with partial structural elements. We focus on structures 

that might arise due to specific metal binding to the unfolded state of 

the protein. There is no assignment of denatured SOD1 available and 

therefore the first step has been to assign the peaks seen in 15 N-HSQC 

spectra of monomeric pseudo-wildtype SOD1 (C6A/C111A, F50E/ 

G51E) in 9 M urea. Experiments performed include both proton and 

carbon detected triple-resonance experiments such as HNCA, HN(CO) 

CA, HNCACB, CANCO and CACO. We have also done chemical shift 

mapping experiments with and without metal addition to conclude 

which amino acids that interact specifically with the metal ions. 

Bi66 

Membrane and peptide interaction of the proinsulin C-peptide 

studied by biophysical methods 

Unnerståle, Sofia; Lind, Jesper; Mäler, Lena 

Stockholm University, Department of Biochemistry and Biophysics, 


The proinsulin C-peptide is produced together with insulin upon 

hydrolysis of specific peptide bonds in the precursor proinsulin. 

Therefore, the role of the C-peptide as a spacer sequence, that 

facilitates the insulin folding process, has been acknowledged. However, 

more recent findings suggest that the C-peptide has hormonal functions 

of its own. For example, the C-peptide is shown to interact with the 

plasma membrane and with insulin. As standard procedure insulin is 

given to diabetes type-1 patients. Nevertheless, a combination of insulin 

and C-peptide might be a better treatment. 

In this study the interaction of the C-peptide and different model 

systems, i.e. membrane mimicking systems and peptides, is studied by 

biophysical methods. Thus, basic biophysical properties of the C-peptide 

can be deduced revealing, for instance, which amino acids that interact 

with the membrane, and which constitution the interacting parts of 

the membrane should have. This study shows that the pH affects the 

structure of the C-peptide, which in turn affects the interaction with 

membrane mimicking systems. 

CD spectroscopy measurements show that the C-peptide is 

predominately unstructured when dissolved in sodium phosphate buffer 

at pH 3.2, 5.8 and at pH 6.9. CD spectra of the C-peptide in POPC and 

POPC/POPG 8:2 also show random coil features at pH 3.2, 5.8 and 6.9, 

respectively. 

TOCSY spectra of the C-peptide in these different conditions reveal that 

the structure is not affected by addition of DMPC nor DMPC/DMPG 9:1 

in pH 5.8. Interestingly, diffusion NMR measurements show that the 

C-peptide interacts with DMPC and DMPC/DMPG, respectively in pH 

5.8. The structure rearrangement, as a consequence of the pH, makes 

the C-peptide prone to interact with bicelles. Furthermore, the effects of 

the C-peptide on different peptides have been studied by the means of 

NMR. Altogether, these results provide a platform for deriving detailed 

features of the C-peptide. 


Bi67 

Towards the oligomeric structure of the chaperone alpha-B 

crystallin - a small heat shock protein 

Markovic, Stefan 1 ; Jehle, Stefan 1 ; van Rossum, Barth-Jan 1 ; Rehbein, 

Kristina 1 ; Bardiaux, Benjamin 1 ; Kühne, Ronald 1 ; Diehl, Annette 1 ; 

Rajagopal, Ponni 2 ; Klevit, Rachel 2 ; Oschkinat, Hartmut 1 

1 FMP Berlin, Berlin, Germany; 2 University of Washington, Seattle, United 

States 

Alpha-B crystallin belongs to the group of small heat shock proteins 

and is predominantly located in the human eye. It’s purpose is to keep 

aggreagation prone proteins in solution. Loss of its chaperone activity 

leads to several diseases including cartaract and desmin-relatedmyopathy. 

As the protein oligomerizes to a polydisperse oligomer of around 30 

monomers, application of X-ray crystallography or solution nuclear 

magnetic resonance spectroscopy (NMR) for structural studies has 

been limited. Using solid-state NMR, the structure of the alpha-B 

crystallin domain in oligomers has been solved previously. In this study, 

intermolecular contacts are measured via different nitrogen-carbon 

correlation methods, such as PAIN, TEDOR and NHHC. A structure of the 

alpha-B crystallin dimer in oligomers is presented. Furthermore, models 

for interactions of higher oligomerization based on intermolecular 

contacts are proposed. Spectra at decreased pH show shift differences 

in these interaction sites suggesting disruption of the oligomer to 

expose the chaperone binding sites. 

Bi68 

Introduction of a paramagnetic tag for NMR conformational 

analysis of oligosaccharides 

Yamaguchi, Takumi 1 ; Yamamoto, Masahiro 2 ; Yamamoto, Sayoko 2 ; Yagi, 

Hirokazu 2 ; Erdelyi, Mate 3 ; Griesinger, Christian 4 ; Kato, Koichi 1 

1 Institute for Molecular Science, Okazaki, Japan; 2 Nagoya City University, 

Graduate School of Pharmaceutical Sciences, Nagoya, Japan; 

3 University of Gothenburg, Department of Chemistry, Göteborg, Sweden; 

4 Max-Planck Institute for Biophysical Chemistry, Göttingen, Germany 

Development of various methods in structural biology has made 

possible to provide three-dimensional structures of biological 

macromolecules. However, conformational characterization of 

oligosaccharides is still challenging. Although NMR spectroscopy 

has great potential for dealing with oligosaccharides which have 

flexible properties precluding X-ray crystallographic approaches, NMR 

analyses of oligosaccharides are frequently hampered by the severe 

spectral overlap resulting from small variations of functional groups of 

glycans and by the insufficiency of conformational restraints based on 

NOE and scalar couplings. Hence, development of methods for NMR 

conformational analyses of oligosaccharides in solution would be highly 

desirable. For its realization, we attempted to introduce a paramagnetic 

tag into oligosaccharides, generating additional distance and angular 

restraints by using paramagnetic effects such as paramagnetic 

relaxation enhancement (PRE), pseudocontact shift and residual dipolar 

coupling. 

An EDTA derivative designed to coordinate a paramagnetic metal 

ion was covalently linked in good yield to the reducing end of N, N’diacetylchitobiose, 

which corresponds to the common core part of 

N-glycans. To an aqueous solution of this modified carbohydrate, Mn(II) 

ion was titrated to give a 1:1 complex. The complexation reaction 

resulted in the decrease of peak intensity caused by PRE in NMR 

measurements. In HSQC spectrum, we observed the modification of the 

signal of each proton according to their positions on the molecule, this 

can provide additional structural information. 


Bi69 

Structure and interaction of human papillomavirus E7 

oncoproteins 

Seiboth, Thomas 1 ; Baum, Marina 1 ; Carella, Michela 1 ; Ramachandran, 

Ramadurai 1 ; Leppert, Joerg 1 ; Ohlenschlaeger, Oliver 1 ; Duerst, Matthias 2 ; 

Goerlach, Matthias 1 

1 Leibniz Institute for Age Research, Biomolecular NMR Spectroscopy, 

Jena, Germany; 2 Friedrich Schiller University, Klinik für Frauenheilkunde 

und Geburtshilfe, Jena, Germany 

High risk (HR) human papillomaviruses (HPV) frequently cause human 

cancers, e.g. the cervix carcinoma, whereas low risk (LR) types mainly 

induce benign lesions. The oncoproteins E6 and E7 encoded in the viral 

genome are key players in cell transformation and their presence is 

necessary to maintain a tumorous state. Both proteins act by targeting 

main tumour suppressor pathways (p53 and pRb) and exert their 

function by direct protein-protein interaction. To better understand the 

structural basis of the oncogenic function of E7, we are investigating 

the structure and interaction of E7 proteins from HR as well as from 

LR HPV. This should allow to answer the intriguing question whether or 

not there are general structural properties related to HR versus LR HPV 

derived E7. 

Based upon our recently published 1 structure of a HR HPV45 E7 we 

are in the process to structurally characterise a further HR E7 variant 

(HPV18) and a variant derived from the lower risk type HPV26. At the 

same time, the interaction of these E7 variants with cellular target 

proteins is addressed. Despite a high structural homology they show 

differences in their dynamics, which might contribute to the different 

interaction behaviour towards their cellular target proteins. 

1 O. Ohlenschlaeger, T. Seiboth, H. Zengerling, L. Briese, A. Marchanka, 

R. Ramachandran, M. Baum, M. Korbas, W. Meyer-Klaucke, M. Duerst, 

M. Goerlach (2006). Solution structure of the partially folded high-risk 

HPV 45 oncoprotein E7. Oncogene 25, 5953-5959. 

Bi70 

Interaction of mithramycin analogues with DNA oligomers 

studied by NMR 

Trigo-Mouriño, Pablo 1 ; Navarro-Vázquez, Armando 2 ; Sánchez-Pedregal, 

Víctor M. 1 

1 Universidade de Santiago de Compostela, Química Orgánica, Santiago 

de Compostela, Spain; 2 Universidade de Vigo, Química Orgánica, Vigo, 

Spain 

Some antibiotics of microbial origin possess antitumour activity due to 

their binding to DNA. One class of these compounds is the group of 

derivatives of aureolic acic, that includes chromomycin and mithramycin 

(MTM) among others.[1] These compounds bind preferentially to the 

minor groove of CG-rich regions of the DNA. The oligosaccharide and 

the R side-chains are crucial elements in their interaction with DNA. 

Recently, by using combinatorial biosynthesis strategies,[2] novel 

analogues of mithramycin have been discovered that display better 

therapeutic index than mithramycin itself. We are especially interested 

in those derivatives that differ in the side chain R, such as MTM-SK 

and MTM-SDK.[3] Our goal is to understand how those small changes 

in structure are related to their different affinity for DNA. We are using 

solution NMR to investigate the structure of some of these ligand/DNA 

complexes. We expect that this information may help propose new 

structures with improved binding properties. 

References. 

[1] L.L. Remsing, H.R. Bahadori, G.M. Carbone, E.M. McGuffie, C.V. 

Catapano, J. Rohr, Biochemistry 2003, 42, 8313. 

[2] C. Méndez, J.A. Salas, Trends Biotechnol. 2001, 19, 449. 

ABSTRACTS POSTER


[3] L.L. Remsing, A.M. González, M. Nuer-e-Alam, M.J. Fernández- 

Lozano, A.F. Braña, U. Rix, M.A. Oliveira, C. Méndez, J.A. Salas, J. Rohr, 

J. Am. Chem. Soc. 2003, 125, 5745. 

Bi71 

Expression and characterization of antimicrobial peptide, 

bovine lactophoricin 

Kim, Ji Sun; Park, Tae Joon; Kim, Yongae 

Hankuk Univ. of Foreign Studies, Chemistry, Yongin, Republic of Korea 

The carboxy terminal 113 to 135 region of component-3 of proteose 

peptone (PP3) which is called Lactophoricin (LPcin-I) is a 23-amino acid 

peptide that corresponds to found in bovine milk. It has been reported 

that LPcin-I has antibacterial activity and a cationic amphipathic helical 

structure, but its shorter analogous peptide (LPcin-II), a 17-amino 

acid peptide, corresponding to the 119 to 135 region of PP3 does 

not display antibacterial activity. In fact, it was interestingly known 

that LPcin-I and LPcin-II have similar charge ratios and identical 

hydrophobic/hydrophilic sectors, according to their helical wheel 

projection patterns, and both peptides show cationic amphipathic 

helical folding and interact with membranes. However, it is known that 

only LPcin-I incorporates into planar lipidic bilayers by forming voltagedependent 

channels. In this study, we cloned and expressed the two 

recombinant peptides as fusion proteins inclusion bodies in E. coli in 

order to understand the structure-activity relationship of these peptides. 

Fusion proteins were purified and recombinant LPcin-I and LPcin-II were 

released from fusion by chemical cleavage. Final purifications were 

achieved by preparative reversed-phase HPLC. We obtained several 

tens of milligrams of uniformly and selectively 15N labeled peptides per 

liter of growth, which was sufficient for solid-state NMR spectroscopy. 

Initial structural data were obtained by solution NMR spectroscopy and 

solid-state NMR spectroscopy compared with and without membranelike 

environments. 

Reference 

1. Protein Expression and Purification 65 (2009) 23 

2. Revue bibliographique: la fraction proteose-peptones 69 (1989) 1 

Bi72 

Structure and interaction of ubiquitin-associated domain of 

human fas associated factor 1 

Song, Jinsue 1 ; Park, Joon Kyu 2 ; Lee, Jae-Jin 3 ; Choi, Yun-Seok 4 ; 

Ryu, Kyoung-Seok 4 ; Kim, Jae-Hong 5 ; Lee, Kong-Joo 3 ; Kim, Eunice 

EunKyeong 2 ; Jeon, Young Ho 4 

1 Chungbuk National University, Chungbuk, Republic of Korea; 2 Korea 

Institute of Science and Technology, Seoul, Republic of Korea; 3 Ewha 

Womans University, Seoul, Republic of Korea; 4 Korea Basic Science 

Institute, Chungbuk, Republic of Korea; 5 Korea University, Seoul, 

Republic of Korea 

Fas associated factor 1 (FAF1) is a multi-domain protein that was first 

identified as a member of the Fas death-inducing signaling complex, 

but later found to be involved in various biological processes. Although 

the exact mechanisms are not clear, FAF1 appears to play an important 

role in cancer, asbestos-induced mesotheliomas, and Parkinson’s 

disease. It interacts with poly-ubiquitinated proteins, Hsp70, p97/ 

VCP in addition to the proteins in the Fas-signaling pathway. We have 

determined the crystal structure of the ubiquitin associated domain of 

human FAF1 (hFAF1-UBA) and examined its interaction with ubiquitin 

and ubiquitin-like proteins using NMR. The hFAF1-UBA revealed a 

canonical three-helical bundle, and it selectively binds to mono- and diubiquitin 

(Lys48-linked), but not to SUMO-1 and NEDD8. The interaction 

between the hFAF1-UBA and di-ubiquitin involves the hydrophobic 

58 

interaction accompanied by a transition in the di-ubiquitin conformation. 

These results provide structural insight into the mechanism of 

polyubiquitin recognition by hFAF1-UBA 

Bi73 

NMR study of isolated voltage-sensing domain from KvAP 

K+ channel: detergent screening, secondary structure and 

backbone dynamics 

Shenkarev, Zakhar; Paramonov, Alexander; Lyukmanova, Ekaterina; 

Shingarova, Lyudmila; Arseniev, Alexander 

Semyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, 


The structure and dynamics of isolated voltage-sensing domain of KvAP 

channel from Aeropyrum pernix (VSD) was studied by high-resolution 

NMR. To find optimal experimental conditions several detergent based 

membrane mimetics were screened. The comparison of 15 N-HSQC 

spectra revealed that spatial structure of VSD in the anionic detergents 

(SDS, LMPC, LMPG) is significantly differ from the structure in the 

zwitterionic or weakly-cationic media (DPC, DMPC/DHPC, LDAO), 

although the both types of media provide undistinguishable CD 

spectra (~60% of helicity). Surprisingly, the 15 N-TROSY spectra of VSD 

measured in environment of zwitterionic (DMPC) or anionic (DMPG) 

lipid-protein nanodiscs, involving patches of lipid bilayer, were very 

similar to spectra of domain in zwitterionic detergents, thus indicating 

that zwitterionic detergents preserve “membrane-like” structure of 

VSD. The almost complete backbone resonance assignment and 

partial side-chain assignment of 2 H, 13 C, 15 N-labeled VSD were obtained 

in the DPC/LDAO mixture. Secondary and spatial structures of VSD 

were characterized using secondary chemical shifts and sparse NOE 

contacts. Results indicate that spatial structure of VSD-KvAP in the 

micelles closely resembles structure of the domain in the inactivated/ 

open state of the channel. Obtained data confirm the presence of interhelical 

salt-bridge (Asp62-R133) and the system of hydrogen bonds 

formed by tightly-bound water molecule, which stabilize overall spatial 

structure of the domain. The backbone dynamics of VSD-KvAP was 

studied using 15 N relaxation measurements. The loop regions S1-S2 and 

S2-S3 were found mobile, and the S3-S4 loop (voltage-sensor paddle) 

was found stable at the ps-ns timescale. The fragments of S1, S2, S3 

and S4 helices located in the region of inter-helical contacts (at the level 

of Asp62-R133 salt-bridge) were found subjected to conformational 

exchange fluctuations at the mks-ms timescale. The obtained data 

permit critical reassessment of some voltage-gating models. Probably, 

observed low-amplitude inter-helical motions represent inherent 

property of the VSD and can play pivotal role in voltage-gating. 

Bi74 

Determination of α-chymotrypsin-glyoxal complex structure 

using 18 O differential isotope shifts 

Spink, Edward; Hewage, Chandralal; Malthouse, J.Paul G 

UCD School of Biomolecular and Biomedical Science, Centre for 

Synthesis and Chemical Biology, Dublin, Ireland 

The substrate derived peptide inhibitor Z-Ala-Pro-Phe glyoxal has been 

shown to be a potent inhibitor of α-chymotrypsin (K i = 19nM)[1]. 13 C 

NMR studies using 13 C enriched inhibitors have shown that Z-Ala-Pro- 

Phe glyoxal forms a tetrahedral complex with chymotrypsin that is 

thought to be analogous to the tetrahedral intermediate formed during 

catalysis. However glyoxals have two electrophilic centres that could 

possibly undergo nucleophilic attack by the active hydroxyl nucleophile 

of the serine 195 residue. Therefore it is essential to determine the 

structure of the tetrahedral species formed between glyoxal inhibitors 

and chymotrypsin to assess its suitability as a transition state analogue. 


In this study 18 O differential isotope shifts were used to determine the 

structure of the chymotrypsin-glyoxal species and assess its suitability 

as a transition state analogue [2]. 

1. Spink, E., Cosgrove, S., Rogers, L., Hewage, C., Malthouse, J.P.G. 

(2007) J. Biol. Chem. 282, 7852-7861 

2. Spink, E., Hewage, C. & Malthouse, J. P. G. (2007). Biochemistry, 46, 

12868-12874 

Computational 


Co10 

DANGLE: A Bayesian inferential method for predicting protein 

backbone dihedral angles and secondary structure 

Cheung, Nicole 1 ; Broadhurst, Bill 1 ; Maguire, Mahon 2 ; Stevens, Tim 1 

1 University of Cambridge, Department of Biochemistry, Cambridge, 

United Kingdom; 2 University of Cambridge, Department of Biochemistry, 

Cambridge, United Kingdom 

We have developed a new computer program called DANGLE, which 

predicts protein secondary structure and backbone phi and psi dihedral 

angles solely from amino acid sequence information, experimental 

chemical shift measurements and a database of known protein 

structure and their associated shifts. The approach uses Bayesian 

inferential logic to analyse the likelihood of conformations throughout 

Ramachandran space and to provide dihedral angle constraints that 

bear a realistic relationship to the error in the estimates. It also assigns 

a secondary structure class for each query residue based on database 

fragments that possess similar shifts and sequences. 

The reliability of angle estimations from DANGLE is high in both helix 

and sheet regions of the Ramachandran plot and is not dependent 

on the population differences in the structure database used by the 

program. When tested on a set of 186 proteins, 88 % and 85 % of 

psi and psi angle estimates, respectively, were within 30 degrees of 

reference angles from high-resolution X-ray structures. In addition, 92 

% and 88 % of reference angles lay within the suggested boundary 

ranges. Simple filtering procedures can identify the most “predictable” 

residues, yielding > 92 % of angle estimates accurate to within 30 

degrees of the reference. In contrast to other approaches, more than 

80 % of phi and psi predictions for glycine and pre-proline residues are 

reliable. Furthermore, DANGLE is able to assign 86 % of the secondary 

structure to the correct class. 

Applications to folded and denatured states of proteins will be 

discussed. 

Co11 

Protein dynamics from a network of coupled rotators: dynamics, 

thermodynamics, comparison with fractional brownian 

dynamics 

Salgado, Gilmar 1 ; Pelupessy, Philippe 2 ; Calandrini, Vania 3 ; Dhulesia, 

Anne 4 ; Ouazzani, Jamal 5 ; Lopes, Philippe 5 ; Bodenhausen, Geoffrey 2 ; 

Kneller, Gerald 3 ; Abergel, Daniel 2 

1 Ecole Normale Supérieure, Paris, France; 2 Ecole Normale Supérieure, 

Département de Chimie, Paris, France; 3 CNRS, Centre de Biophysique 

Moléculaire, Orléans, France; 4 Cambridge University, Department of 

Chemistry, Cambridge, United Kingdom; 5 CNRS, Institut de Chimie des 

Substances Naturelles, Gif-sur-Yvette, France 

Spin relaxation measurements offer valuable insight into protein 

dynamics. We have recently introduced a simple NMR-oriented model 

based on a Network of Coupled Rotators (NCR) to describe the internal 

dynamics of proteins. [1] This model allows one to predict 15N relaxation 

rates based on the sole knowledge of the three-dimensional structure. 

[2,3] Using this approach, it is possible to calculate equilibrium quantities, 

such as order parameters [4] . In addition, an analytical relationship 

between the order parameter and conformational entropy can be 

derived. This relationship illustrates the absence of a one-to-one 

relationship between order parameters S 2 and conformational entropy, 

and explains the difficulty to relate S 2 obtained from experimental data 

to the former [5] . 

A radically different approach to analyze NMR relaxation, and based 

ABSTRACTS POSTER


on fractional Brownian dynamics (fBD), was recently introduced. 

Comparison with NCR has shown recently [6] that the correlation 

functions of bond vectors in a protein, as predicted by the NCR model, 

can be interpreted in terms of a heuristic approach based on fractional 

Brownian dynamics for each vector in the network. A comparison of 

NCR and fBD approaches on the calcium-binding protein calbindin will 

be presented. In addition, analysis of 15N relaxation measurements 

performed on the 266-residue protein 6-phosphogluconolactonase 

(6PGL) [7] in terms of the various models will be discussed. 

References 

[1] D. Abergel, G. Bodenhausen, 2005 J. Chem. Phys., 123, 204901 

[2] G. Nodet, G. Bodenhausen, D. Abergel, 2008 C.R. Chimie , 11, 524 

[3] G. Nodet, D. Abergel, G. Bodenhausen, 2008 ChemPhysChem, 9, 

625-633 

[4] A. Dhulesia, D. Abergel, G. Bodenhausen, 2007 J. Am. Chem. Soc., 

129, 4998 

[5] A. Dhulesia, G. Bodenhausen, D. Abergel, 2008 J. Chem. Phys., 129, 

095107 

[6] V. Calandrini, D. Abergel, G. Kneller, 2008 , J. Chem. Phys., 128, 

145102 

[7] M. Delarue et al., 2007 J. Mol. Biol., 366, 868-881 

Co12 

Multiple quantum NMR of spin-carrying molecules of a gas in 

nanopores 

Doronin, Serge; Fedorova, Anna; Fel’dman, Edward; Zenchuk, Alexandre 

Institute of Problems of Chemical Physics, Theoretical Department, 

Chernogolovka, Russian Federation 

It is well known [1,2] that the dipole-dipole interactions of nuclear 

spins of molecules of a gas are not averaged completely by molecular 

diffusion in nanopores. As a result, it is possible to perform multiple 

quantum (MQ) NMR experiments [3] in nanopore compounds. The MQ 

NMR Hamiltonian at the preparation period of the MQ NMR experiment 

[3] commutes with the operator of the square of the total spin angular 

momentum I^2. Thus it is suitable to study MQ NMR dynamics in the 

basis of common eigenstates of I^2 and Iz, where Iz is a projection of 

I^2 on the external magnetic field. Then the problem is reduced to MQ 

NMR dynamics of subsystems with all possible values of the total spin 

angular momentum (I, I-1, I-2,…). Usually one can investigate MQ NMR 

dynamics of not more than fifteen spins even using supercomputer 

calculations [4]. However the introduced basis allows us to avoid 

problems connected with a big number of spins and to investigate MQ 

NMR dynamics of more than 600 spins using a personal computer. We 

have found an analytical solution of MQ NMR dynamics of a system 

consisting of nine spins. Numerical investigations allow us to find exact 

profiles of intensities of MQ coherences and to study the dependencies 

of spin numbers in many-spin clusters which are responsible for MQ 

NMR coherences. 

The work is supported by the Program of the Presidium of Russian 

Academy of Sciences No.27. 

1.J.Baugh, A.Kleinhammes, D.Han, Q.Wang, Y.Wu, Science 294, 1505 

(2001). 

2.E.B.Fel’dman, M.G.Rudavets, JETP 98, 207 (2004). 

3.J.Baum, M.Munovitz, A.N.Garroway, A.Pines. J. Chem. Phys.83, 2015 

(1985). 

4.S.I.Doronin, E.B.Fel’dman, I.Ya.Guinzbourg, I.I.Maximov, Chem. Phys. 

Lett.341, 144 (2001). 

60 

Co13 

Predicting amyloid aggregation rates of proteins using 

multivariate analysis 

Malm, Linus 1 ; Sjöström, Michael 2 ; Antti, Henrik 2 ; Larsson, Göran 1 

1 Umeå University, Department of Medical Biochemistry and Biophysics, 

Umeå, Sweden; 2 Umeå University, Department of Organic Chemistry, 

Umeå, Sweden 

Aggregation of proteins into amyloid fibrils is a process of great 

interest, as amyloid plaque is involved in diseases such as Alzheimer’s, 

Creutzfeldt-Jakob’s, Parkinson’s disease and others. 

In this study we use a large database of amyloid fibrillation kinetic 

data from mutational studies of human muscle acylphosphatase 

and α-synuclein. The database was analyzed using the multivariate 

methods, principal component analysis and orthogonal partial least 

squares. The amino acid sequence of each protein was described by 

two sets of scales, the first describing physical and chemical properties 

of each amino acid side chain and the second describing the propensity 

of each amino acid to form different types of secondary structures. 

The calculation of the effect on each specific site of mutation was 

complemented by a pattern analysis, performed by auto- and crosscovariance. 

Using these methods we are able predict the effect of 

each mutation on the amyloid aggregation rate of each protein. Our 

method also allow us to describe what changes in psychochemical 

and secondary structure characteristics that are of importance for the 

differences in aggregation rates between the wild type and mutated 

proteins in our database. 

Co14 

Microstructure of hydration shells of CH COO 3 - 2- and SO ions by 

4 

NMR-relaxation and quantum-chemical methods 

Vovk, Mikhail; Pavlova, Maria; Chizhik, Vladimir 

Saint-Petersburg State University, Department of Quantum Magnetic 

Phenomena, Saint-Petersburg, Russian Federation 

The NMR-relaxation method is one of the most fruitful techniques for 

the investigation of systems consisting of different substructures with 

fast molecular exchange among them like in electrolyte solutions. 

The work is devoted to the investigation of the microstructure of 

aqueous solutions containing CH COO 3 - 2- and SO ions. Some important 

4 

parameters of the nearest vicinity of the ions, such as ion coordination 

numbers and quadrupole coupling constants (QCC) of deuterons of 

the water molecules belonging to the first hydration shells, have been 

estimated using NMR-relaxation method. 

The quantum-chemical calculations have been used to simulate 

hydration shells of the ions and to interprite the experimental data. 

The molecular-ionic clusters CH COO 3 - (D O) (where n varies in a wide 

2 n 

2- range: from 4 to 24 water molecules) and SO (D2O) (n = 8 and 24) 

4 n 

have been considered as models of substructures in the solutions. 

For example, 24 water molecules form the hydration shell of the 

2- SO ion with 8 and 16 water molecules in the first and second layers, 

4 

consequently. The geometry of clusters were optimised at B3LYP/6- 

31++G** level of theory. The distribution of water molecules near 

hydrophobic and hydrophilic ion sides of the CH COO 3 - ion was obtained 

2- and it was compared with experimental results. For the SO ion the 

4 

QCC values were calculated for deuterons of water molecules belonging 

to the first hydration shell. 

A noticable decrease in the QCC values obtained in the relaxation 

experiments and in the calculations allowed the investigation of possible 

2- hydrogen-bond structures around the SO ion and their influence on 

4 

QCC values. 

The work is supported by the RFBR grant # 07-03-00735a 


Co15 

The generating functions formalism for the analysis of spin 

response to the periodic trains of RF pulses 

Lukzen, Nikita; Petrova, Marina; Koptyug, Igor; Savelov, Andrei; Sagdeev, 

Renad 

International Tomography Center SB RAS, Novosibirsk, Russian 

Federation 

Long trains of periodic RF pulses make an integral part of MRI 

methods. Extensively used multiple echoes can serve as an example 

of such kind. We present a new effective approach for calculation of 

magnetization evolution under the influence of trains of periodic RF 

pulses in the framework of the generating functions (GF) formalism. 

Generating function is defined as a function of complex variable: 

F(z)=M 1 +M 2 z+M 3 z 2 +...+M n z n +... Here, is, for instance, the n-th echo 

amplitude. Generating function comprises complete information about 

all echo amplitudes at once. 

In this work it is shown that in many cases closed analytical expression 

for the generating function can be found. From GF, the magnetization at 

the step (period) with arbitrary number n can then be easily calculated. 

The generating functions approach is especially efficient for analysis 

of multiecho sequences, when to find the echo amplitude one has 

to average over different isochromates. Earlier in our paper [1] we 

obtained analytical expressions for GF and echo amplitudes in Carr- 

Purcell-Meiboom-Gill pulse sequence with an arbitrary refocusing angle. 

In present work we generalized this GF approach for calculation of echo 

amplitudes for CPMG sequence of selective pulses for an arbitrary 

resonance offset and RF magnetic field magnitude and also for gradient 

echo sequence. Comparison of calculated and experimental echo 

amplitudes has been done for refocusing pulse equal to ϖ/4. 

[1]. N. Lukzen, A.A. Savelov, Analytical derivation of multiple spin echo 

amplitudes with arbitrary refocusing angle, J. Magn. Reson. 2007, 185, 

71-76. 

Co16 

Modeling Xe chemical shifts due to interaction with single-wall 

carbon nanotubes 

Lantto, Perttu 1 ; Vaara, Juha 2 

1 University of Oulu, Department of Physical Sciences, Oulu, Finland; 

2 University of Helsinki, Department of Chemistry, Helsinki, Finland 

Xenon guests are used intensively in NMR spectroscopy as “spy” atoms 

in various materials due to its inertness and highly sensitive NMR 

chemical shift. It provides detailed experimental information of material 

properties of e.g. porous solids [1]. 

Theoretical description of Xe chemical shift inside fullerenes is shown 

to necessitate inclusion of both relativistic and dynamic effects [2]. The 

recently developed Breit-Pauli perturbation theory (BPPT) [3] of leadingorder 

relativistic effects on nuclear shielding enables the inclusion 

of electron correlation effects on the Xe shift. This is so far the only 

method by which ab initio electron-correlated relativistic shieldings 

are available. By using BPPT, the correlation effects, especially in the 

nonrelativistic part of the shielding tensor, have been found to constitute 

a challenge for the present DFT methods in xenon compounds such as 

xenon fluorides [4] and the novel HXeCCH molecule [5]. Quantitative 

theoretical description of the experimentally probably best-characterized 

intermolecular interaction effect in NMR, the temperature dependence 

of the second virial coefficient of 129 Xe shielding, was reached by 

including both relativistic and high-level electron correlation effects in 

the binary Xe chemical shift function [6]. 

In the present work, previous experience [2-6] is exploited in the 

theoretical description of Xe NMR shift in single-wall carbon nanotubes 

(SWCNT). The dependence of relativistic Xe shift on the adsorption site 

both inside and outside the nanotubes is settled. Also the sensitivity of 

the Xe shift to the SWCNT type is explored. 

[1] J. Jokisaari, Encyclopedia of Spectroscopy and Spectrometry (J. C. 

Lindon, G. E. Tranter, and J. L. Holmes, Academic, New York, 1999). 

[2] M. Straka, P. Lantto, and J. Vaara, J. Phys.Chem. A. 112, 2658 

(2008). 

[3] P. Manninen, K. Ruud, P. Lantto, and J. Vaara, J. Chem. Phys. 122, 

114107, (2005), (E) 124, 149901, (2006). 

[4] P. Lantto and J. Vaara, J. Chem. Phys. 127, 084312, (2007). 

[5] M. Straka, P. Lantto, M. Räsänen, and J. Vaara, J. Chem. Phys. 127, 

234314, (2007). 

[6] M. Hanni, P. Lantto, M. Ilias, H. J. Aa. Jensen, and J. Vaara, J. Chem. 

Phys., 127, 164313, (2007). 


Co17 

Determination of native ensembles using RDCs with explicit 

tensor estimation 

Esteban-Martín, Santiago; Fenwick, Bryn; Salvatella, Xavier 

Institute for Research in Biomedicine (IRB Barcelona), Laboratory of 

Molecular Biophysics, Barcelona, Spain 

Residual dipolar couplings (RDC) are a well established tool for both the 

refinement of biomolecular structures and for the elucidation of their 

underlying dynamics in the sub-ms time scale. The interpretation of 

RDCs in multi-domain systems and partially folded states is however 

challenging as current state-of-the-art methods rely on the fitting of 

ensembles of conformations to a single alignment tensor or on the 

use of RDCs for the validation of pre-defined ensembles. Here, we 

expand the applicability of RDCs to both multi-domain and intrinsically 

disordered proteins by using ensemble molecular dynamics simulations 

restrained by RDCs with an explicit calculation of the alignment tensor. 

We show that RDCs can indeed be used to reconstruct distributions of 

conformations for multi-domain and partially folded proteins without a 

priori assumptions 

Co18 

From NMR data to structure deposition: The Extend-NMR 

software pipeline 

Stevens, Timothy 1 ; Boucher, Wayne 1 ; Fogh, Rasmus 1 ; Steiner, Peter- 

René 2 ; Guigas, Bruno 2 ; Jaravine, Victor 3 ; Orekhov, Vladislav 4 ; Staykova, 

Doroteya 4 ; Billeter, Martin 4 ; Brunner, Konrad 5 ; Kalbitzer, Hans Robert 5 ; 

Rieping, Wolfgang 1 ; Lamazhapova, Darima 1 ; Bardiaux, Benjamin 6 ; 

Nilges, Michael 6 ; van Dijk, Marc 7 ; Bonvin, Alexandre 7 ; Doreleijers, 

Jurgen 8 ; Vuister, Geerten 8 ; Vriend, Gert 8 ; Penkett, Chris 9 ; Vranken, Wim 9 ; 

Henrick, Kim 9 ; Laue, Ernest 1 

1 Unversity of Cambridge, Department of Biochemistry, Cambridge, 

United Kingdom; 2 Bruker BioSpin GmbH, Karlsruhe, Germany; 3 Goethe 

University Frankfurt am Main, Frankfurt am Main, Germany; 4 University 

of Gothenburg, Gothenburg, Sweden; 5 University of Regensburg, 

Institute of Biophysics and Physical Biochemistry, Regensburg, 

Germany; 6 Institut Pasteur, Paris, France; 7 Utrecht University, Bijvoet 

Center for Biomolecular Research, Utrecht, Netherlands; 8 Radboud 

University, Nijmegen, Netherlands; 9 European Bioinformatics Institute, 

Hinxton, Cambridge, United Kingdom 

The Extend-NMR project is a collaboration between European NMR 

software developers who have collectively developed an NMR software 

suite to support functional and structural proteomics. The Extend-NMR 

software pipeline encompasses the NMR software process from data 

acquisition and processing through resonance assignment, analysis and 

structure determination to validation and finally database deposition. 

All of the software elements in the Extend-NMR pathway are linked via a 

ABSTRACTS POSTER


single graphical user interface and all of the open-source elements are 

installable as a single computational package. The pipeline is designed 

to eliminate the difficulties that a user may have when moving between 

different software tasks, and enables the system to be used as a highthoughput 

tool for structural and functional genomics. 

The individual elements of the Extend-NMR pipeline are as follows: 

TopSpin (Bruker BioSpin GmbH); Data acquisition and processing, 

MDD (Orekhov); Multiway decomposition processing of nonuniformly 

sampled spectra, PRODECOMP (Billeter); Processing highdimensionality 

projection spectra, AUREMOL (Kalbitzer); Bayesian 

peak picking, CcpNmr (Laue/CCPN); NMR data pipeline, assignment 

software and data format conversion, ARIA (Nilges); Automated NOE 

assignment and structure calculation, ISD (Nilges/Rieping); Bayesian 

inferential structure calculation, HADDOCK (Bonvin); High-ambiguity 

macromolecular docking, CING (Vuister/Vriend); Macromolecular 

structure validation, Deposition (Henrick/PDBe); Streamlined deposition 

to PDB and BMRB, 

The tight connection between all of the software components in the 

Extend-NMR project has been made possible by using the CCPN 

software development infrastructure. CCPN provides a unifying platform 

for all NMR software by providing a comprehensive data model and 

corresponding computer libraries, in multiple programming languages, 

which describe all aspects of the NMR data pathway. 

Co19 

Configurational analysis of dibromo-palauàmine using DFT 

calculations 

Reinscheid, Uwe 1 ; Köck, Matthias 2 ; Griesinger, Christian 1 

1 MPIBPC, NMR II, Göttingen, Germany; 2 Alfred-Wegener-Institut, 

Bremerhaven, Germany 

In case of flexible molecules in solution, the determination of the 

relative and absolute configuration is a formidable task, which can 

be ambiguous even with the modern arsenal of analytical techniques. 

NMR spectroscopy constitutes the most important method to determine 

configuration and conformation in solution. The available chemical 

shift information can be used to establish the relative configuration 

by comparison with DFT (density functional theory) calculated values. 

If optical rotation and/or VCD (vibrational circular dichroism) data 

are available, even the absolute configuration can be determined in 

favourable cases. The combined application (chemical shift, optical 

rotation and VCD prediction by DFT calculations) to a controversial 

stereochemical problem will be presented: the stereochemistry of the 

family of palauàmines for which a reassignment was recently proposed 

(Grube et al., 2007). 

Grube A, Köck M (2007) Structural assignment of tetrabromostyloguanidine: 

Does the relative configuration of the palauàmines need 

revison ?, Angew. Chem. Int. Ed., 46, 2320 

Co20 

Multithreaded simulation of Dnmr spectra on cuda enabled 

GPGPU video cards 

Szalay, Zsófia; Rohonczy, János 

Eötvös Loránd University, Budapest, Hungary 

Computer simulations based on the Monte Carlo method are time 

consuming calculations. The spreading of multicore processors 

gave the possibility to reduce runtime by dividing the calculation on 

multiple CPUs. Even faster calculations can be performed on GPGPU 

(General Purpose Graphic Processor Unit) based NVidia video cards 

implementing the CUDA programming language extension. Reports on 

very fast simulations can be found in the literature based on this parallel 

62 

multiprocessor architecture [1] but these methods have not appeared 

on the field of NMR spectrum simulations yet. 

The dynamic NMR spectrum simulation would require huge computer 

RAM and long calculation time if the common solution of the 

Liouville-von Neumann equation would be applied. Even if memory 

reducement techniques like symmetry adoption [2] and efficient matrix 

diagonalization algorithms [3] are used, more than seven spins per 

conformer cannot be simulated without strong simplifications. 

An alternative theory is based on the Monte Carlo simulation. DNMR 

spectra are calculated by averaging the spectra of individual molecules 

[4-6]. We have applied this technique to simulate dynamic NMR spectra 

of large spin systems with multisite exchanges. Our program has 

been extended by using the GPGPU hardware and CUDA technique. 

According to our extensive tests it performs the simulation ten times 

faster than the original one running on CPU. 

References 

[1] M. S. Friedrichs et al., J. Comp. Chem. 2009 DOI: 10.1002/ 

jcc.21209 

[2] G. Binsch, J. Am Chem. Soc. 1969 91 1304 

[3] R. S. Dumont, S. Jain, A. Bain, J. Chem. Phys. 1997 14 106 

[4] J. K. M. Sanders, B. K. Hunders, Modern NMR Spectroscopy, Oxford 

Univ. Press 1987. 

[5] Zs. Szalay, J. Rohonczy, J. Magn. Reson. 2008 191 56 

[6] Zs. Szalay, J. Rohonczy, J. Magn. Reson. 2009 197 48 

Co21 

A graphical interface for the analysis of quantum mechanically 

computed NMR properties 

Navarro-Vázquez, Armando 

Universidade de Vigo, Departamento de Química Orgánica, Vigo, Spain 

Computation of NMR properties, such as chemical shielding tensors 

or scalar coupling constants, by quantum mechanical methods is 

gaining momentum in the field of NMR based structural determination 

of organic compounds. Most popular quantum chemical programs can 

compute most important properties such as chemical shielding tensors, 

scalar coupling constants and magnetic susceptibilities at different 

levels of theory. We present here a new module for the NMRDev 

platform ( http://webs.uvigo.es/webqo3/people/armando/software/ 

nmrdev ) which intends to help spectroscopists and chemists in the 

analysis of NMR QM computations. 

This new module can parse computed properties from different program 

packages such as Gaussian03, GAMESS or NwChem. Chemical shifts 

can be automatically calculated form chemical shielding tensors if 

a suitable reference is provided. The program allows also graphical 

representation of the principal axes of the tensors. If scalar coupling 

constants are computed the program allows extraction of different sets 

according to selected bonding patterns. Last but not least, magnetic 

susceptibility tensors can be also extracted and graphically represented. 

Program is freely available under open source license. 

Co22 

Simultaneous multi-way decomposition of projections from 

different experiments 

Fredriksson, Jonas; Doroteya, Staykova; Billeter, Martin 

Gothenburg University, Biophysics, Gothenburg, Sweden 

Multi-Way Decomposition of projections of high-dimensional spectra 

offers a variety of possibilities for comprehensive characterisations of 

proteins: complete resonance assignment, 3D structure and more. By 

combining different projection experiments for the same protein we are 

able to resolve backbone assignment and NOE information. Additional 


projection experiments can then give side-chain information completing 

the description of the protein. 

When the directly detected dimensions of two otherwise different 

experiments coincide, then the corresponding projections can be 

merged into one data set. Since decompositions require that the signals 

of each projection fit into the corresponding spin system, the merging of 

two 5D experiments provide unambiguous 9D spin systems. A selection 

of 5D and 4D experiments, for example Hα/β-Cα/β-C’-N-NH and Hα/β- 

Cα/β-N-C’-N-NH, that are centred on the same H-N moiety can be used 

for backbone characterization. The second experiment exhibits lower 

sensitivity and therefore can be replaced by experiments where either 

NOEs or TCOSY-steps represent the central magnetisation transfer. 

At the same time the TOCSY- and NOESY-based experiments provide 

information for side chain assignments and 3D structure, respectively. 

All projections are two-dimensional and have the chemical shift of HN 

along the directly detected dimension. The other dimension corresponds 

to a linear combination of several chemical shifts, including at least 

one shift of N or C’ combined with a varying selection of shifts from the 

α and β carbons and hydrogens of both residues. The joint multi-way 

decomposition of all spectra gives descriptions of spin systems, each 

consist of all nuclei in a CβHn–CαH-C’-NH-CαH-CβHn fragment that 

stretches over two adjacent residues. 

1 Staykova, D.K.; Fredriksson, J., Bermel, W., Billeter, M. J. Biomol. NMR 

2008, 42, 87-97. 

Co23 

“Fast NMR” or “Accurate NMR”? Towards new applications of 

random sampling 

Kazimierczuk, Krzysztof; Zawadzka, Anna; Kozminski, Wiktor 

University of Warsaw, Chemistry, Warsaw, Poland 

Level of artifacts in spectra obtained by Multidimensional Fourier 

Transform has been studied, considering randomly sampled signals 

of high dimensionality and long evolution times. It has been shown 

theoretically and experimentally, that this level is dependent on the 

number of time domain samples, but not on its relation to the number of 

points required in appropriate conventional experiment. Independence 

of the evolution time domain size (in the terms of both: dimensionality 

and evolution time reached), suggests that random sampling should 

be used rather to design new techniques with large time domain than 

to accelerate standard experiments. This conclusion is in contardiction 

with common way of employing of random sampling in “Fast NMR”. It 

seems to be more proper to use it in “Accurate NMR” i.e. in experiments 

allowing precise determination of spectra parameters because of 

high resolution and high dimensionality. 5D HC(CC-TOCSY)CONH 

has been presented as the example of such approach. The feature of 

Multidimensional Fourier Transform, namely the possibility of calculating 

spectral values at arbitrary chosen frequency points, allowed easy 

examination of resulting spectrum. We present the example of such 

approach, referred to as Sparse Multidimensional Fourier Transform. 


Co24 

Co-Processing NMR spectra with multi-dimensional 

decomposition 

Orekhov, Vladislav 1 ; Hiller, Sebastian 2 ; Samuelsson, Linda 3 ; Jaravine, 

Victor 1 ; Ibraghimov, Ilghiz 4 ; Larsson, Joakim 3 ; Wagner, Gerhard 2 

1 University of Gothenburg, Swedish NMR Centre, Gothenburg, Sweden; 

2 Harvard Medical School, Department Biological Chemistry and 

Molecular Phar, Boston, United States; 3 University of Gothenburg, 

Department of Physiology, Gothenburg, Sweden; 4 Elegant Mathematics 

Ltd, Ottweiler, Germany 

Analysis of typical sets of multidimensional NMR experiments for signal 

assignment, structure calculations, and metabolomic studies, relies 

on matching of signal frequencies between several experiments. By 

simultaneous processing of the spectra, this intrinsic reoccurrence of 

signal frequencies and line-shapes is exploited to enhance quality and 

efficiency of the data analysis. We use multi-dimensional decomposition 

(MDD) for co-processing of any combination of experiments and nonuniform 

sampling for optimizing spectra resolution and sensitivity. The 

approach has been successfully demonstrated for several essentially 

different situations: processing of 4D NOESY spectra for the de novo 

structure determination of the 31 kDa integral human membrane 

protein VDAC-1 in detergent micelles with effective molecular weight 

of 70-90 kDa; rapid real-time data collection and automated backbone 

assignments of 13 kDa naturally disordered cytoplasmic part of the 

zeta-chain of the T-cell receptor system; a metabolomic study on fooddeprived 

fish. 

References: 

1. Hiller, S., R.G. Garces, T.J. Malia, V.Y. Orekhov, M. Colombini, and G. 

Wagner, Science, 2008, 321, 1206-10. 

2. Jaravine, V. A. and Orekhov, V. Y. J Am Chem Soc 2006, 128, 13421-6. 

Co25 

NMRDepot: New database for experimental NMR spectra 

Szalay, Zsófia; Rohonczy-Boksay, Erzsébet; Rohonczy, János 

Eötvös Loránd University, Budapest, Hungary 

Anyone who has started studying a new group of chemicals has met the 

problem of finding suitable reference spectra for well known standard 

molecules. There are NMR spectrum databases on the internet as well 

as books of spectrum collections [1]. Most of these cannot be updated 

easily and are collected by a group of scientists resulting in a rather 

selected database. 

There are also free databases of NMR chemical shifts on the internet 

those can be updated and edited easily and the subject of the spectra 

are not restricted to the area of interest of the authors [2,3]. However a 

database for chemical shifts does not give the same (full) information as 

the measured spectra itself. 

A new website, NMRDepot is introduced now which handles a database 

of full 1D and 2D experimental spectra of various molecules [4]. The 

server is written in Java language and it is based on the Apache’s 

Tomcat program and a MySQL database. Spectrum displaying, 

downloading and uploading are managed by Java Applets. 

The NMRDepot database can be searched for various spectrum 

parameters. The hits are displayed as small previews of the spectra with 

a few parameters. A detailed page of each spectrum is also available 

by displaying it in an interactive applet thus enabling online zooming 

and scaling of the spectrum. There is an option of downloading the 

spectrum in Bruker format and viewing it in any spectrum processing 

program. 

Registered users can upload their spectra and the most important 

parameters are extracted from the spectrum file itself. The user only 

has to specify the compound’s name, its chemical formula and the 

ABSTRACTS POSTER


spectrum directory to upload the spectrum. We hope that the easiness 

of uploading will encourage everyone to share their spectra with the 

NMR community. 

References 

[1] SDBSWeb : http://riodb01.ibase.aist.go.jp/sdbs/ 

[2] NMRShiftDB: http://nmrshiftdb.ice.mpg.de/ 

[3] nmrdb.org: http://www.nmrdb.org 

[4] NMRDepot: nmrdepot.chem.elte.hu 

Co26 

Automated assignment of solid-state NMR spectra using 

GARANT 

Carlsson, Jonas 

University of Gothenburg, Department of Chemistry, Göteborg, Sweden 

A novel method for automated assignment of solid-state nuclear 

magnetic resonance (ssNMR) protein spectra is presented and 

illustrated on the small immunoprotein GB1. 

ssNMR presents a novel situation compared to solution NMR since 

often no 1 H information is used. We use the existing assignment 

program GARANT, which was designed for solution NMR data. Hence, 

magnetization transfers for various ssNMR spectra had to be defined in 

GARANT before any assignment trials could be performed. 

Four different ssNMR spectra were first manually peak picked (both 2D 

and 3D ones). GARANT discriminates between nuclei of the same type 

but with different shift ranges, e.g. between carboxyl and alpha carbons. 

Thus, peak lists from e.g. a CC spectrum could be split up into lists with 

or without carboxyl carbons, according to the normal shift intervals for 

these atom types based on statistics from BMRB, resulting in a total of 

eight peak lists. 

It was found that highly complete assignments were indeed achievable. 

In the backbone of the protein, only 5 out of 56 amide nitrogen atoms 

are not assigned and none erroneously. The corresponding numbers 

for the 56 C atoms are 3 and 1, respectively, and for the 56 carboxyl 

carbons 3 and 0. All in all, the backbone assignments are 99.4% 

correct and 93.4% complete. 

64 

Enhhanced NMR 

En10 

Fast small excitation angle two-dimensional spectra with in situ 

DNP-NMR 

Ludwig, Christian; Marin-Montesinos, Ildefonso; Saunders, Martin; 

Günther, Ulrich 

University of Birmingham, School of Cancer Sciences, Birmingham, 


Dynamic nuclear polarisation (DNP) is used to transfer the high spin 

polarisation of unpaired electrons to coupled nuclear spins by doping 

samples with a stable radical and irradiating with microwaves at the 

EPR lines of the radical. Signal enhancements of ~200 were achieved 

in the solid state [1,2] whereas enhancements ~10,000 were reported 

after polarising the sample at low temperature (1.4K), followed by 

dissolution with hot solvent and transfer of the sample to an NMR 

spectrometer [3]. 

While this implementation of DNP (Hypersense) achieves a much 

higher enhancement than possible at room temperature it only 

allows a single shot acquisition of the NMR spectrum because of the 

dissolution and transfer of the sample to the high resolution NMR 

magnet. Therefore acquisition of multidimensional NMR spectra 

must be achieved within the relaxation time of the hyperpolarisation. 

Recently the applicability of ultrafast 2D-NMR acquisition has been 

demonstrated in conjunction with DNP-NMR [4]. This method is very 

demanding in terms of spectrometer hardware and massive use of 

gradients causes a loss of intensity. Here we present an alternative 

approach to acquire 2D-NMR spectra in a single run by using only a 

fraction of the polarisation in a single fid through a small flip-angle 

excitation. Acquisition of a 2D-spectrum in a single run eliminates 

errors introduced by variations between multiple polarisations. Several 

possibilities were explored to create a small flip-angle excitation. 

This sequence has been demonstrated for several small molecule 

pharmaceuticals and reduced acquisition times from 5-6h to 3-5sec. 

References 

[1] DA Hall, DC Maus, GJ Gerfen, SJ Inati, LR Becerra, FW Dahlquist, RG 

Griffin. Science 276, 930-932 (1997) 

[2] CT Farrar, DA Hall, GJ Gerfen, M Rosay, J-H Andenkjær-Larsen, RG 

Griffin. J. Magn. Reson. 144, 134-141 (2000) 

[3] JH Ardenkjær-Larsen, B Fridlund, A Gram, G Hansson, L Hansson, 

MH Lerche, R Servin, M Thaning, and K Golman. Proc. Nat. Acad. Sci. 

USA, 100, 10158-10163 (2003) 

[4] L. Frydman and D. Blazina; “Ultrafast Two-Dimensional Nuclear 

Magnetic Resonance Spectroscopy of Hyperpolarized Solutions”; 

submitted 

En11 

Biocompatible, spin-labeled Heparins as polarizing agents for 

dynamic nuclear polarization (DNP) 

Hinderberger, Dariush 1 ; Dollmann, Björn 1 ; Kleshyov, Andrey 2 ; Schreiber, 

Laura 2 ; Spiess, Hans 1 ; Münnemann, Kerstin 1 

1 Max Planck Institute for Polymer Research, Mainz, Germany; 2 University 

of Mainz, Mainz, Germany 

A versatile and biocompatible class of spin-labeled macromolecules 

was investigated by continuous wave (CW) electron paramagnetic 

resonance (EPR), double electron-electron resonance (DEER) and 

room-temperature dynamic nuclear polarization (DNP) that can be 

utilized for in vivo magnetic resonance imaging (MRI) and EPR imaging 

(EPRI). All presented heparin radicals show reasonably high 1 H DNP 


enhancement factors up to E = -91 despite their very broad EPR lines. 

The distributions of the spin labels were scrutinized and compared 

with the crystallographic structure of heparin. Remarkably, the heparin 

radicals with dipolar coupling frequencies (from EPR/DEER analysis) 

matching the proton Larmor frequency manifested the best DNP-NMR 

enhancements. The heparin nitroxides may also be suitable for an 

efficient hyperpolarization of 13 C containing molecules and for low 

temperature DNP measurements due to their broad range of dipolar 

coupling frequencies. Furthermore, they can be applied themselves as 

hyperpolarized and functional substances that bind to the endothelium. 

En12 

Spectral properties of hyperpolorized xenon detected without rf 

excitation: maser emissions and nuclear spin-noise detection 

Desvaux, Hervé; Marion, Denis J.Y.; Huber, Gaspard; Berthault, Patrick 

CEA, Gif sur Yvette, France 

In liquid-state NMR, resorting to dissolved xenon, polarized at a high 

level (>20%) thanks to spin-exchange optical pumping induces the 

appearance of new physical phenomena due to distant dipolar fields 

(DDF) and non-linear coupling between the magnetization and the 

rf coil (radiation damping). As a consequence a reassessment of 

conclusions drawn for thermal equilibrium systems is required and new 

perspectives appear. We shall here report our latest results obtained 

by simply monitoring the NMR signal without coherent rf excitation of 

the xenon magnetization. When negative xenon spin temperature is 

selected during the optical pumping step, the analysis of the multiple 

spontaneous maser emissions indicate a linear correlation between 

their radiative energy and a rate characteristic of their life-time, tending 

to validate the key importance of DDF in this chaotic behavior [1]. On 

the other hand, for positive xenon spin temperature the first detection 

of nuclear spin-noise of an hyperpolarized species is reported. We show 

that conversely to situations encountered for thermal equilibrium, this 

approach gives directly access to the spectral and dynamic properties 

of Xenon. In fact, the conditions of detection are fully renewed, making 

this approach particularly promising for very small number of spins, 

even at low static magnetic field. This concept is validated by spin-noise 

detection of a number of spin smaller than what it can be detected by 

one pulse experiment at thermal equilibrium [2]. 

[1] D.J.Y. Marion, P. Berthault, H. Desvaux, “Spectral and temporal 

features of multiple spontaneous NMR-maser emissions”, Eur. Phys. J. 

D 51 (2009) 357-367. 

[2] H. Desvaux, D.J.Y. Marion, G. Huber, P. Berthault, “First nuclear 

spin-noise spectra of hyperpolarized systems”, Angewandte Chemie, In 

press. 

En13 

Performance of an integrated dissolution DNP spectrometer for 

liquid state NMR spectroscopy 

Leggett, James; Panek, Rafal; Granwehr, Josef; Perez-Linde, Angel; van 

der Drift, Anniek; Köckenberger, Walter 

University of Nottingham, SPMMRC, School of Physics and Astronomy, 

Nottingham, United Kingdom 

Recently it was demonstrated that large nuclear spin polarisation 

in liquid state samples can be generated by first increasing the 

polarisation of the nuclear spin system in solid state using dynamic 

nuclear polarisation (DNP) at low temperature followed by a fast 

dissolution step [1]. 

A two-centre, integrated 3.4 T DNP polariser and 9.4 T liquid state NMR 

spectrometer has previously been presented [2]. Due to the proximity 

of the two magnetic centres in such a system, the polarised sample 

can be rapidly transferred and, furthermore, this can be done in the 

solid state followed by subsequent dissolution immediately above the 

NMR centre. This significantly reduces T1 relaxation loss, as well as 

eliminating cross-relaxation that can arise when liquid-state samples 

are shuttled through a varying magnetic field. Consequently it is 

possible to observe signals from very short T1 species in both natural 

abundance 13C spectroscopy and low concentration 1H spectroscopy. 

Here we present results obtained using small peptide samples. The 

acquired liquid state enhancements depend exponentially on the T1 

values. For the proton groups used, the T1 value varies between 0.4 

and 2.5 s. A comparison of the enhancement factor, †, is provided 

between the integrated system and a system consisting of a standalone 

polariser connected to a separate 9.4T magnet. In natural 

abundance 13C spectroscopy experiments using the stand-alone 

polariser, it was found that 3 of the 8 Ala-Gln di-peptide carbon 

resonance frequencies could not be observed due to the short T1 of 

these lines. However, using the dual-centre system it was possible 

for all resonance lines to be detected with significant enhancement. It 

is found that fast shuttling of the sample in the solid state allows for 

maximal exploitation of the DNP enhancement. 

References 

[1] Ardenkjaer-Larsen et al, PNAS, 100(18) p10436-10439 (2003) 

[2] Leggett et al, EUROMAR 2008, Poster EMR-18 

Acknowledgements - We are very grateful for support from G Smith 

and R Hunter from the University of St Andrews, Scotland, UK. The 

project was funded by an EPSRC instrument development grant and a 

technology development grant from the BBSRC. 


En14 

A 129 Xe-biosensor for monitoring MHC:peptide interactions 

Mitschang, Lorenz 1 ; Kilian, Wolfgang 1 ; Schlundt, Andreas 2 ; Sticht, 

Jana 2 ; Freund, Christian 2 ; Guenther, Sebastian 3 ; Hoepner, Sabine 3 ; Falk, 

Kirsten 3 ; Roetzschke, Olaf 3 

1 Physikalisch-Technische Bundesanstalt, Medical Physics, Berlin, 

Germany; 2 Leibniz-Institute of Molecular Pharmacology, Protein 

Engineering Group, Berlin, Germany; 3 Max-Delbrueck-Center for 

Molecular Medicine, Berlin, Germany 

Progress in biosensor design depends on the development of sensitive 

probes for the recognition of molecular processes. Because of its 

unique properties, laserpolarized 129Xe can yield molecule-specific 

MR spectroscopic and imaging information with high sensitivity. Based 

on the work of the Pines group, a novel 129Xe-biosensor is presented 

which enables the detection of complex formation of a peptide ligand 

with Major Histocompatibility Complex class II (MHC II). MHC II are 

cellular surface molecules which trigger T cell-mediated immune 

responses when bound to an antigenic peptide molecule. 

The biosensor consists of a crytophane cage functionalized by binding 

through an amino-PEG-acid linker the hemagglutinin (HA) peptide 

(N-terminally extended by GEEG). The polar linker and the charged 

peptide yield an excellent solubility of the construct, whereas HA 

ensures high specificity (affinity 100 – 300 nM) of complex formation 

because of strong binding with MHC II (allel Human Leukocyte Antigen 

DR1). Also the cage-linker moiety does not alter the binding mode of HA 

to MHC II, as is revealed by essentially identical 15N-HSQC spectra of 

such a complex in comparison to a complex of wildtype HA with MHC II. 

Finally, the functional integrity of MHC II in complex with the construct is 

evident from T-cell activation in an in-vitro assay. 

NMR detection of complex formation relies on the accessibility of the 

crytophane cage to solute xenon atoms in slow exchange: besides the 

bulk signal at 194 ppm, a second line is detected at 64.5 ppm for the 

construct alone in solution which, however, is shifted 1 ppm downfield 

for the construct bound to MHC II. Experiments are performed with 

10 uM construct/MHC II and 8 mM 129Xe laserpolarized to 15% in 2 

ml PBS with a home-built 1H/129Xe probe on a 3T whole body MRI 

ABSTRACTS POSTER


scanner. A further gain in sensitivity of 2 to 3 orders is achieved upon 

using chemical exchange saturation transfer. 

The solubility, specificity, MR-sensitivity and structural and functional 

integrity of the 129Xe-biosensor for MHC II peptide interactions may 

bear the potential for molecular imaging studies in-vitro and in-vivo of 

immune response and autoimmune disease. 

En15 

13 C Hyperpolarization studies of unsaturated substrates by 

using Parahydrogen Induced Polarization and appropriate pulse 

sequences 

Roth, Meike 1 ; Bargon, Joachim 2 ; Koch, Achim 1 ; Spiess, Hans W. 1 ; 

Münnemann, Kerstin 1 


of Bonn, Bonn, Germany 

Parahydrogen induced polarization has turned out to be a versatile 

technique to obtain hyperpolarized molecules exhibiting strong NMR 

signals via a chemical approach. PHIP makes use of breaking the high 

initial symmetry of parahydrogen during homogeneously catalyzed 

hydrogenations of unsaturated substrates. Consequently, the population 

of the energy levels of their spin states deviates from the Boltzmann 

distribution characteristic for systems in thermal equilibrium. This 

leads to absorption and emission signals in the NMR spectra and a 

theoretical signal increase of up to 10 5 , which is in practice limited, 

however, by relaxation processes in the product. Transfer of polarization 

to hetero-nuclei can be implemented randomly in weak magnetic fields 

or selectively via special pulse sequences. Optimization of polarization 

transfer to e.g. 13 C is crucial for applications like metabolic imaging 

where the highest possible 13 C polarization is required to obtain high 

SNR images [1]. 

To achieve a polarization transfer to 13 C we applied the PH-INEPT and 

the PH-INEPT+ sequence to a PASADENA experiment under pressure at 

elevated temperature [2, 3]. Regarding our model compound 1-hexyne 

the PH-INEPT+ sequence showed higher signal enhancements. The 

obtained 13 C PH-INEPT+ NMR spectra display polarization transfer to all 

carbons of the hydrogenation product 1-hexene. Corresponding to the 

chosen delay the polarization can be transferred selective to different 

carbon nuclei. 

Spontaneous polarization transfer from 1 H to 13 C in the 

parahydrogenation product 1-hexene under ALTADENA conditions only 

showed signal enhancements less than 100. By applying appropriate 

pulse sequences to a “PASADENA under pressure” experiment we 

achieved polarization transfers to 13 C yielding a signal enhancement of 

up to 4000. 

[1] P. Battacharya, E. Y. Chekmenev, W. H. Perman, K. C. Harris, A. P. 

Lin, V. A. Norton, C. T. Tan, B. D. Ross, D. P. Weitekamp, J. Magn. Reson. 

186, 150-155 (2007). 

[2] M. Haake, J. Natterer, J. Bargon, J. Am. Chem. Soc. 118, 8688- 

8691 (1996) 

[3] M. Roth, J. Bargon, H. W. Spiess, A. Koch, Magn. Reson. Chem. 46, 

713-717 (2008) 

66 

En16 

Generation of 13 C hyperpolarized barbituric acid derivatives via 

Parahydrogen Induced Polarization 

Roth, Meike 1 ; Bargon, Joachim 2 ; Spiess, Hans W. 1 ; Koch, Achim 1 ; 

Münnemann, Kerstin 1 


of Bonn, Bonn, Germany 

The application of 13 C NMR spectroscopy and imaging for clinical 

diagnostics has been constrained by the extremely long acquisition 

times that are required to obtain high SNR under physiological 

conditions. However, this obstacle could be overcome by in vitro 

hyperpolarisation of a molecule with long 13 C spin lattice relaxation time 

via Parahydrogen Induced Polarization (PHIP) and subsequent injection 

into the animal or patient of investigation. Hence, the role of certain 

target compounds such as anesthetics could be investigated by using 

MRI techniques. Among the drugs used to treat epilepsy, barbiturates 

like 5-methyl-5-propargylbarbituric acid are attractive from the MRI and 

biomedical point of view. 

5-Methyl-5-propargylbarbituric acid was synthesized from urea and 

methyl-propargylmalonic acid. The unsaturated group is used to 

introduce polarization into the molecule according to standard PHIP 

procedures [1]. Parahydrogen was generated by cooling thermal 

hydrogen with a closed-cycle cryostat setup in the presence of active 

charcoal to achieve an enrichment of up to 98%. In order to enhance 

the conversion rate of the hydrogenation reaction of 5-methyl-5propargylbarbituric 

acid, the PASADENA experiment (chemical reaction 

inside the spectrometer) is carried out at elevated temperature and 

pressure [2]. The high proton polarization is transferred to 13 C using a 

PH-INEPT+ sequence with different delays [3]. 

It was shown before by our group that homogeneous hydrogenation 

of unsaturated barbituric acid derivatives with 50% parahydrogen 

yielded a substantial increase of the 1 H-NMR signals of the reaction 

products. However, signal enhancement by randomly triggered 

polarization transfer to 13 C in the weak magnetic field could not be 

observed. Application of a closed-cycle cryostat setup for parahydrogen 

enrichment up to 98% together with effective INEPT-derived pulse 

sequences allowed for 13 C NMR signal enhancements up to 2200. 

[1] J. Natterer, J. Bargon, Prog. Nucl. Magn. Reson. Spectrosc. 31, 293 

(1997) 

[2] M. Roth, J. Bargon, H. W. Spiess, A. Koch, Magn. Reson. Chem. 46, 

713- 717 (2008) 

[3] M. Haake, J. Natterer, J. Bargon, J. Am. Chem. Soc. 118, 8688- 

8691 (1996) 

En17 

Hyperopolarized gas-phase MR imaging of reactions in 

microreactors 

Anwar, Muhammad Sabieh 1 ; Bouchard, Louis-S. 2 ; Burt, Scott R. 3 ; 

Kovtunov, Kirill V. 4 ; Koptyug, Igor V. 4 ; Pines, Alexander 3 

1 School of Science and Engineering, Lahore University of Management 

Sciences (LUMS), Lahore, Pakistan; 2 University of California at Los 

Angeles, Department of Chemistry, Los Angeles, United States; 

3 University of California at Berkeley, Department of Chemistry, Berkeley, 

United States; 4 International Tomography Centre, Institutskaya St., 

Novosibirsk, Russian Federation 

We demonstrate magnetic resonance imaging (MRI) in the gas phase 

using para-hydrogen (p-H 2 ) induced polarization. A reactant mixture 

of H 2 enriched in the para spin state and propylene gas is flowed 

through a reactor cell containing a heterogenized catalyst, Wilkinson’s 

catalyst immobilized on modified silica gel. The hydrogenation product, 

propane gas, is transferred to the NMR magnet and is spin-polarized 


due to the ALTADENA effect. A polarization enhancement factor of 300 

over thermally polarized gas was observed in 1D proton spectra. The 

enhancement was also evident in the magnetic resonance images of 

phantoms placed inside an NMR tube. This is the first demonstration of 

imaging a hyperpolarized gaseous product formed in a hydrogenation 

reaction catalyzed by a supported catalyst. This result may lead to 

several important applications, including flow through porous materials, 

gas-phase reaction kinetics and adsorption studies and MRI in low 

fields, all using catalyst-free polarized fluids, and that too, in the gas 

phase. For example, we use this approach to demonstrate the mapping 

of chemical reactions in a microreactor. We can also spatio-temporally 

control the delivery of the polarized product through pulse sequence 

engineering. Using the appropriate pulse sequences, we can lock 

the polarization state of the polarized product, enhance its lifetime 

and finally “release” the polarization as it reaches remote parts of the 

microreactor. 

En18 

Principle of operation and performance of a “Triplet DNP” 

polarizer 

Jannin, S. 1 ; Haag, M. 2 ; Hautle, P. 2 ; Konter, J.A. 2 ; van den Brandt, B. 2 ; 

Ansermet, J.-Ph. 3 ; Wenckebach, W. Th. 4 ; Comment, A. 5 ; van der Klink, 

J.J. 5 

1 EPFL, IPN and LRMB, Lausanne, Switzerland; 2 PSI, SEPT, Villigen, 

Switzerland; 3 EPFL, IPN, Lausanne, Switzerland; 4 PSI and EPFL, SEPT 

and IPN, Lausanne, Switzerland; 5 EPFL, IPN and CIBM, Lausanne, 


Dynamic Nuclear Polarization (DNP) has become a practicable 

technique to enhance sensitivity in NMR. Free mono- or bi- radicals are 

used to enhance the nuclear spin polarization by factors of up to 300. 

Taking advantage of the high electron spin polarization of the photoexcited 

triplet state of pentacene, we developed an X-band “Triplet 

DNP” polarizer (operating in a field of 3 kG), and performed DNP in 

a pentacene doped naphthalene glassy sample. This technique goes 

far beyond the theoretical proton spin polarization enhancement limit 

provided by conventional DNP methods with free radicals and does 

not require either sophisticated cryogenic equipment or high field. 

In fact, the electron spin polarization available, after photo-excitation 

of the pentacene, is temperature and field independent and exceeds 

80% (maximum theoretical DNP enhancement E max =250 000). At 

the operating conditions of our apparatus, at a field of 3 kG and at 

a temperature of 100 K, the thermal equilibrium polarization of a 

conventional free radical electron spin is only 0.2% (E max =660). Another 

great advantage of this method is that nuclear spin lattice relaxation 

is practically not affected by the presence of the polarizing agents, as 

is the case with standard free-radical doped samples. With the “Triplet 

DNP” approach, one can tune the polarizing agent concentration 

down to zero by simply switching off the excitation light. In the late 

eighties, W.Th. Wenckebach et al. [1] invented the technique using 

single crystals, and more recently, M. Takeda et al. [2] showed it is also 

quite effective on powder ground from a crystal. In this contribution, 

we show that this approach also works well in samples obtained by 

rapidly cooling a melt (a “glass”) and we describe some features of 

our apparatus. Glassy samples could eventually host molecules of 

interest. We thus show that the scope of application of the method can 

be broadened and that “Triplet DNP” has the potential to become an 

interesting tool in solid state NMR and chemistry. 

[1] Chem. Phys. Lett. 165(1):6-10, 1990 

[2] Chem. Phys. Lett. 345(1):166-170, 2001 


En19 

Structure determination of large protein complexes and 

transient intermediates by solution NMR techniques 

Madl, Tobias 1 ; Simon, Bernd 2 ; Mackereth, Cameron 3 ; Gabel, Frank 4 ; 

Nilges, Michael 5 ; Sattler, Michael 1 

1 Helmholtz Zentrum and Technische Universitaet Muenchen, Institute of 

Structural Biology, Garching, Germany; 2 EMBL Heidelberg, Heidelberg, 

Germany; 3 Institut Européen de Chimie et Biologie, Bordeaux, France; 

4 Institut Laue Langevin, Grenoble, France; 5 Institut Pasteur, Paris, France 

The functions of biomacromolecules depend on their molecular 

interactions and assemblies in larger multimeric complexes which 

play important roles e.g. in the regulation of gene expression and/or 

cellular signaling. While the structural characterization of the individual 

independent structural units is fairly standard, there is a lack of efficient 

methods for a detailed analysis of the quaternary arrangement of the 

domains in a complex. Furthermore, complex formation often involves 

transient intermediates, requiring the use of solution techniques to 

study these complexes. 

We have developed efficient strategies for structural analysis of 

complexes and transient intermediates by combining residual dipolar 

couplings (RDCs), paramagnetic relaxation enhancements (PREs) from 

spin labels and inert paramagnetic co-solvents (solvent PREs), as well 

as small angle X-ray/neutron scattering data (SAXS/SANS). Binding 

interfaces between the individual subunits are mapped based on 

chemical shift perturbations, NOEs, and solvent PREs. Relative domain 

orientation between the subunits is constrained with by RDCs, PREs 

and SAS. We have implemented a flexible and robust protocol in the 

molecular dynamics/simulated annealing program CNS/ARIA that allows 

structure calculation of the quaternary arrangement of the complexes 

by direct refinement against the experimental data. 

Our approach is demonstrated on a multi-protein/RNA complex that 

defines the 3’ intron-exon boundary at the early stages of the tightly 

controlled pre-mRNA (alternative) splicing pathway in humans. We show 

that prior recognition of the 3’ splice site, the splicing factor U2AF65 

subunit pre-exists in distinct conformations. Intriguingly, we found 

that one of these conformers resembles the preformed RNA-bound 

arrangement. Following the principle of a conformational selection 

mechanism, U2AF65 undergoes a substantial population shift upon 

RNA-binding and leads to a single RNA-bound conformation. The 

exchange between these conformations is modulated by the sequence 

of the intron RNA, such that the population shift correlates with RNA 

binding affinity and splicing. 

En20 

Exploring new radicals for solution DNP applications 

Gabellieri, Cristina 1 ; Mugnaini, Veronica 2 ; Oliveros, Malena 2 ; Feliz, 

Miguel 3 ; Veciana, Jaume 2 ; Pons, Miquel 4 

1 Institute for Research in Biomedicine (IRB), Barcelona, Spain; 2 Institut 

de Ciencia de Materials (CSIC) and CIBER-BBN, Bellaterra, Spain; 

3 University of Barcelona, Bercelona, Spain; 4 Institute for Research in 

Biomedicine (IRB) and University of Barcelona, Barcelona, Spain 

Dynamic Nuclear Polarization (DNP) is attracting considerable attention 

as a method to increase the NMR sensitivity in selected applications. 

Non equilibrium nuclear polarization is transferred from electron spin 

polarization by microwave irradiation at frequencies corresponding 

to electronic transitions (including hyperfine couplings). This is most 

efficiently carried out in the solid state at low temperatures. For slowly 

relaxing nuclei, non-equilibrium polarized samples can be transferred to 

a conventional NMR spectrometer and studied in solution. 

The choice of radical is crucial for the success of the experiment. 

Factors like the width of the EPR line in the solid state compared 

with the nuclear frequency determine the mechanism of polarization 

ABSTRACTS POSTER


transfer. Additional effects can arise from the different chemical nature 

of the radical used, including solubility and supramolecular interactions 

between the molecule to be polarized and the free radical. 

We have studied a new series of trityl radicals not previously used 

for DNP experiments. The new radical tested vary in their substitution 

leading to different symmetry properties. In all cases we could 

demonstrate DNP effects. For some of these radicals, we see striking 

differences in the sign of the nuclear polarization generated in different 

molecular targets, emphasizing the role of supramolecular interactions 

in DNP. 

En21 

Application of parahydrogen-induced nuclear spin polarization 

for the determination of heterogeneous hydrogenation reaction 

mechanism 

Zhivonitko, Vladimir 1 ; Kovtunov, Kirill 1 ; Koptyug, Igor 1 ; Beck, Irene 2 ; 

Bukhtiyarov, Valery 2 

1 International Tomography Center SB RAS, Novosibirsk, Russian 

Federation; 2 Boreskov Institute of Catalysis SB RAS, Novosibirsk, 


It is well-known that involvement of parahydrogen into the 

hydrogenations lead to strong enhancement of NMR signal intensity if 

reaction occur in pairwise manner with the formation of nonsymmetrical 

products. This phenomenon called parahydrogen-induced polarization 

(PHIP) was widely used for the investigations of homogeneous 

hydrogenation reactions, but for a long time it was under the question 

that observation of PHIP is possible for heterogeneous systems. 

Recently, this question was resolved and it is shown that PHIP can be 

observed even in hydrogenations over metal supported heterogeneous 

catalysts [1,2]. In the present work we considered the use of PHIP for 

the derivation of important features of heterogeneous hydrogenation 

mechanism. Propene and propyne hydrogenations were used as model 

reactions. Wide rage of Pt and Pd metal catalyst supported on the ZrO 2 , 

TiO 2 , SiO 2 and Al 2 O 3 was considered. The particles size was varied from 

12 nm to 1 nm and below. Distinguished metal particle size effects and 

support influence on PHIP intensity and polarized NMR signal forms 

were detected. Moreover, stereoselectivity of parahydrogen molecule 

addition during heterogeneous hydrogenations was studied. 

Acknowledgements 

Authors are grateful for the support of the present work by grants 

from the RFBR (08-03-00661, 08-03-00539, 08-03-91102, 07-03- 

12147), the program of support of leading scientific schools (NSh- 

3604.2008.3), CRDF (RUC1-2915-NO07), RAS (5.1.1) and SB RAS 

integration grants (67, 88). 

References 

[1] K.V. Kovtunov, I.E. Beck, V.I. Bukhtiyarov, I.V. Koptyug, Angew. Chem. 

Int. Ed., 2008, 47, 1492-1495. 

[2] K.V. Kovtunov, I.V. Koptyug, “Parahydrogen-induced polarization 

in heterogeneous catalytic hydrogenations”, in: Magnetic Resonance 

Microscopy. Spatially Resolved NMR Techniques and Applications (Codd, 

S.Seymour, J.D., eds), 2008, pp. 101-115. 

68 

En22 

High-field dynamic nuclear polarization in aqueous solutions of 

various radicals 

Gafurov, Marat; Denysenkov, Vasyl; Prandolini, Mark; Endeward, 

Burkhard; Lyubenova, Sevdalina; Prisner, Thomas 

Institut für Physikalische und Theoretische Chemie and Center for 

Biomolecular Magnetic Resonance, Goethe-University Frankfur, 

Frankfurt am Main, Germany 

Large dynamic nuclear polarization (DNP) enhancements of liquid-state 

high-field NMR opens up the possibility of overcoming the current NMR 

sensitivity limits and allows the study of macromolecules complexes 

under physiological low concentrations. Our approach is to polarize 

liquid samples in-situ at high magnetic fields using a double-resonance 

structure at both NMR and microwave EPR frequencies [1]. 

The microwave side of the resonant structure has two important 

features: firstly, it drastically reduces the microwave electrical field 

strength at the sample position, thus avoiding excessive heating of the 

liquid sample; and secondly it strongly enhances the MW magnetic 

field strength at the sample position, which allows significant DNP 

enhancements already with a very low incident MW power of less than 

45 mW. 

Using this system, unexpected high DNP enhancements of more than 

10 have been achieved in liquid water samples at room temperature 

and magnetic fields of 9.2 T (corresponding to 400 MHz 1H NMR 

frequency and 260 GHz EPR frequency) [2]. In this work we compare 

the DNP efficiencies of various organic and inorganic radicals: 

Nitroxides, Fremy’s Salt, and Trityl in aqueous solutions at 9.2 T. These 

results demonstrate the first important step towards the application of 

DNP to high-resolution NMR. 

[1] V.P. Denysenkov, M.J. Prandolini, A. Krahn, M. Gafurov, B. Endeward, 

T.F. Prisner, Appl. Magn. Reson. 2008, 34, 289 

[2] M.J. Prandolini, V.P. Denysenkov, M. Gafurov, B. Endeward, T.F. 

Prisner, J. Am. Chem. Soc. 2009, 131 (17), 6090 

En23 

A liquid-state shuttle DNP spectrometer for 600MHz NMR: 

Construction and results for 1 H and 13 C signal enhancement 

Reese, Marcel 1 ; Türke, Maria-Teresa 2 ; Igor, Tkach 2 ; Marquardsen, 

Thorsten 3 ; Tavernier, Andreas 3 ; Höfer, Peter 3 ; Engelke, Frank 3 ; Bennati, 

Marina 2 ; Griesinger, Christian 1 

1 Max Planck Institute for Biophysical Chemistry, NMR based Structural 

Biology, Göttingen, Germany; 2 Max Planck Institute for Biophysical 

Chemistry, Electron Paramagnetic Resonance, Göttingen, Germany; 

3 Bruker Biospin, Karlsruhe, Germany 

Following the goal of sensitivity enhancement of high resolution liquid 

state NMR we have developed a field cycling by sample shuttling liquid 

state DNP spectrometer (Fig. 1). We polarize the sample at low field 

(0.34T, LF), allowing for relatively large sample diameter and volume 

with reduced heating, and then pneumatically transfer the sample within 

120ms to the high field (14T, HF) for high resolution and high sensitivity 

NMR detection. 

Compared to the 14T Boltzmann signal, enhancement factors of ε HF = 

/ =-3.4 for water protons and ε=+8.4 for carbon in 13 C 

labelled urea (Fig. 1) have been achieved. In experiments using signal 

accumulation of several scans these enhancements translate into a 

reduction of measurement time of a factor of 12 and 70, respectively. 

13 C shuttle DNP experiments with the radical dissolved in organic 

solvents, namely 13 C labelled chloroform and tetrachloromethane, yield 

the enhancements ε HF =+21 and ε HF =+9.2, respectively. 

The highest achieved LF enhancement factors for the protons of water 

are ε LF = / =-170 using 25mM TEMPONE- 15 N- 2 D. The 13 C 


HF enhancement ε HF =+8.4 translates to positive LF enhancements ε 

LF >+336 using 4M 13 C-Urea in D 2 O containing 50mM TEMPONE- 15 N- 2 D. 

For the latter 20s microwave irradiation with 22W has been applied. The 

temperature increase was measured to be 10K using the chemical shift 

temperature dependence of residual H 2 O. To our knowledge this carbon 

enhancement is the so far highest achieved for 13 C nuclei of molecules 

dissolved and polarized in liquid water close to room temperature. 

Surprisingly we have also observed large negative 13 C enhancements 

under very similar conditions. The reason for this change of sign is 

under investigation now. 

Molecular size dependent losses of magnetization during sample 

transfer through low stray fields (≥5mT) of the magnets hinder the 

achievement of large enhancements on large molecules like proteins. 

To allow for enhanced measurements of proteins we are currently 

constructing a dual centre magnet system to shorten the transfer 

distance and transfer time and to assure high magnetic fields on the 

whole sample track. 

En24 

1H and 13C Dynamic Nuclear Polarization for High Resolution 

NMR in Liquid Solutions 

Türke, Maria-Teresa 1 ; Reese, Marcel 1 ; Tkach, Igor 1 ; Marquardsen, 

Thorsten 2 ; Tavernier, Andreas 2 ; Höfer, Peter 2 ; Engelke, Frank 2 ; 

Griesinger, Christian 1 ; Bennati, Marina 1 

1 Max Planck Institute for Biophysical Chemistry, Göttingen, Germany; 

2 Bruker, Rheinstetten, Germany 

Dynamic nuclear polarization (DNP) provides a powerful tool to 

enhance the sensitivity of NMR by transferring the larger electron spin 

polarization to nuclei of interest. In liquid state, DNP is governed by the 

Overhauser mechanism which rapidly loses efficiency with increasing 

magnetic field. One possibility to use low field DNP for high resolution 

NMR is to polarize at 0.34 T (9.6 GHz electron pumping frequency) and 

subsequently shuttle the sample into a 14 T magnet for NMR detection 

(600 MHz 1H / 150 MHz 13C) [1]. 

To test this concept we have extensively investigated the mechanism 

of polarization transfer at 0.34 T in aqueous solution [2,3]. To 

optimize the pumping conditions we have set up a separate low field 

DNP spectrometer. It is based on a Bruker ELEXSYS X-band EPR 

spectrometer and a Bruker minispec (15 MHz 1H) coupled to an 

ENDOR cavity. An option for nitrogen gas cooling of the sample has 

been implemented. With this setup large 1H signal enhancements up to 

ε(0.34T) = / = -170 on water samples containing 15N-2H- 

TEMPONE as polarizer have been achieved. Saturation studies have 

been performed and Overhauser parameters have been evaluated. 

With the prototype of a shuttle DNP spectrometer 1H water 

enhancements up to ε(14T) = -3.4 have been observed which 

translates into ε(0.34T) = (14T/0.34T) * ε(14T) = -140 close to the 

value observed with the low-field setup. The discrepancy can be 

explained by differences in instrumentation and relaxation losses 

during sample transfer. 13C shuttle DNP experiments with the radical 

dissolved in 13C labelled chloroform and tetrachloromethane yield 

ε(14T) = +21 and ε(14T) = +9.2, respectively. As a first test towards 

DNP on biological samples the 13C enhancement on urea dissolved 

in aqueous solution alongside the radical was measured to be -5.7 to 

+8.4. Currently, a dual center magnet is being constructed to minimize 

relaxation losses on the pathway and thus allow high field NMR 

enhancements on aqueous protein samples. 

[1] M. Reese et al., Appl. Magn. Reson., 2008, 34, 301-311 

[2] P. Höfer et al., J. Am. Chem. Soc., 2008, 130, 3254-3255 

[3] P. Höfer et al., Appl. Magn. Reson., 2008, 34, 393-398 


En25 

Photoinitiated radical polymerizations: Reversible addition and 

disproportionation reactions, investigated by CIDNP 

Griesser, Markus; Neshchadin, Dmytro; Gescheidt, Georg 

Graz University of Technology, Institute of Physical and Theoretical 

Chemistry, Graz, Austria 

Radical polymerization has been an important technological and 

scientific field for many years. More recent developments involve 

“living procedures” which depend on reversible addition/elimination 

steps. However, in the framework of “classical” radical polymerization 

reversible steps are not well established. 

The focus of our work is the initial phase of radical polymerization which 

is decisive for its efficiency and the properties of the polymer chain. 

Interesting aspects are the way the initiating radical reacts and followup 

reactions not contributing to chain growth. 

We have performed a mechanistic investigation of the early steps 

of photoinitiated radical polymerization using magnetic-resonance 

techniques (photo-Chemically Induced Dynamic Nuclear Polarization 

(photo-CIDNP)) and theoretical calculations. 

In the course of these experiments we observed that the initial addition 

of vinyl monomers (acrylates, styrene) to a radical may occur reversibly. 

Furthermore we could identify a disproportionation mechanism which, 

amongst other products, generates benzaldehyde, an unwanted 

byproduct in photoinitiated radical polymerizations. 

References: 

M. Griesser, D. Neshchadin, K. Dietliker, N. Moszner, R. Liska, G. 

Gescheidt, 2009, submitted 

K. Dietliker, S. Broillet, B. Hellrung, P. Rzadek, G. Rist, J. Wirz, D. 

Neshchadin, G. Gescheidt, Helv. Chim. Acta 2006, 89, 2211 

D. Hristova, I. Gatlik, G. Rist, K. Dietliker, J.-P. Wolf, J.-L. Birbaum, A. 

Savitsky, K. Moebius, G. Gescheidt, Macromolecules 2005, 38, 7714 

En26 

Biomolecular in vitro applications of solution DNP-NMR 

Meier, Sebastian 1 ; Karlsson, Magnus 2 ; Jensen, Pernille R. 2 ; Baumann, 

Herbert 3 ; Ardenkjær-Larsen, Jan H. 4 ; Duus, Jens Ø. 1 ; Lerche, Mathilde 

H. 2 

1 Carlsberg Laboratory, Valby, Denmark; 2 Albeda Research, Valby, 

Denmark; 3 GE Healthcare, Uppsala, Sweden; 4 GE Healthcare, Hillerød, 

Denmark 

DNP-NMR has experienced a renaissance in the form of a dissolution 

protocol, which provides sensitivity gains up to >10 4 for solution NMR. 

This sensitivity enhancement has mainly been exploited for in vivo tracer 

experiments of metabolism and tissue pH. 

Here, we show that biomolecular solution NMR can benefit significantly 

from the vastly improved detection limit in DNP-NMR. As an 

example, the detection of otherwise elusive reaction intermediates at 

submicromolar concentrations is shown in single-scan 13 C NMR spectra 

of real-time enzymatic assays. Such direct detection of enzymatic 

intermediates has the potential to delineate multistep reaction 

mechanisms. The potentially different relaxation behaviour of reactants 

in different organ compartments (blood, interstitium, intracellular) poses 

a challenge for quantitative in vivo NMR. This obstacle is obviously 

not faced in vitro, which permits a precise quantitative analysis of the 

enzyme kinetics. 

We further show that 13 C DNP-NMR bears great potential for the 

detection of molecular interactions. The 13 C nucleus has a substantially 

larger chemical shift dispersion than the proton and is a more generally 

applicable screening nucleus than 19 F. 13 C DNP-NMR screening 

experiments with 200 µM ligands at natural 13 C abundance or 2 µM 

isotope enriched ligand are shown to provide ample signal for the 

ABSTRACTS POSTER


screening of protein-ligand interactions. 

In summary, 13 C DNP-NMR in solution bears a great promise for 

overcoming sensitivity limitations of a variety of solution NMR 

applications and allows new sets of experiments to be devised for 

academic and industrial applications. 

En27 

The diagonal-free 3D/4D H,N-TROSY-NOESY-TROSY-N,H 

experiment 

Diercks, Tammo 

CiC bioGUNE, NMR platform, Derio, Spain 

The (SQC-) TROSY technique bases on spin state selectivity (S 3 ) to 

prevent mixing of I ± S α and I ± S β single-quantum coherences (SQC), 

allowing to keep apart the TROSY coherence with reduced transverse 

relaxation. This increases resolution, but discards the complementary 

50% anti-TROSY magnetisation. While this loss is unfortunate for 

sensitivity-limited NOESY experiments, their spectra may be even more 

limited by overlap and dynamic range problems caused primarily by the 

intense diagonal signals. The S 3 principle underlying TROSY then offers 

a critical added benefit by allowing to remove diagonal signals through 

orthogonal spin state selection (oS 3 ), since diagonal I ± S α (or I ± S β ) 

polarisation largely conserves its S spin state, while NOE transfer to I’ z 

= ½[I’ z S α +I’ z S β ] looses any S 3 . Applying oS 3 before and after NOE mixing 

then allows only the 50% NOE cross-signal polarisation with altered S 

spin state to pass, blocking both diagonal and the 50% NOE intensity 

with same S spin state. 

Published diagonal-free implementations of the H,N-TROSY-NOESY-H 

experiment (1-2) prepare initial S 3 polarisation H z N α/β by spin-stateselective 

1 H excitation (S 3 E), or by a conventional H,N-TROSY module. 

After NOE mixing, all employ another S 3 E module to directly read out 

observable H - N β TROSY coherence, yielding diagonal-free 2D H,H S 3 E- 

NOESY-S 3 E or 3D H,N-TROSY-NOESY-S 3 E spectra, respectively. 

We illustrate how diagonal removal by oS 3 can be extended to (n≤4) 

D H,N-TROSY-NOESY-TROSY-N,H experiments. The novelty lies in also 

using S 3 transfer after the NOE mixing, rather than a simple INEPT 

module that levels out any S 3 and, thus, allows for passage of 50% 

diagonal signals. For this, we adopt the modified ST2-PT module 

presented in the qTROSY scheme (3) to quantitatively transfer selected 

H z N α cross-peak polarisation into the subsequent second TROSY 

module. As an illustration, we show the diagonal-free 3D [H]N-TROSY- 

NOESY-TROSY-N,H spectrum of MBP that resolves several critical H N -H N 

contacts with near-degenerate 15 N shifts. 

1) Zhu, G. et al J Biomol NMR 1999, 14, 377-381 

2) Meissner, A.; Sorensen, O. W. J Magn Reson 2000, 142, 195-8 

3) Diercks, T.; Orekhov, V. Y. J Biomol NMR 2005, 32, 113-27 

En28 

Structural and dynamic properties of defect dipoles in 

piezoelectric materials 

Erdem, E. 1 ; Erünal, E. 1 ; Drahus, M. D. 1 ; Jakes, P. 1 ; Kiraz, K. 2 ; Somer, M. 2 ; 

Smyth, D. M. 3 ; Zhang, L. X. 4 ; Ren, X. 4 ; Eichel, R.-A. 1 

1 Albert-Ludwig University of Freiburg, Institute of Physical Chemistry, 

Freiburg, Germany; 2 University of Koc, Department of Chemistry, 

Istanbul, Turkey; 3 Lehigh University, Materials Science and Engineering, 

Betlehem, United States; 4 Xián Jiaotong University, Multi-Disciplinary 

Materials Research Center, Xián, China 

The defect structure of aliovalent transition-metal and rare-earth 

functional centers in ferroelectric perovskite oxides is characterized 

by means of multifrequency electron paramagnetic resonance 

spectroscopy, assisted by density-functional theory calculations. The 

70 

review is mainly focused on lead titanate PbTiO 3 compounds. The 

results include the formation of charged defect dipoles, causing internal 

bias fields, multivalence manganese centers, acceptor-type copper 

functional centers creating isolated oxygen vacancies that promote 

ionic conductivity. Moreover, the impact of the defect structure on 

macroscopic material properties is discussed. The effect of external 

electric fields on the orientation of defect dipoles in ferroelectric 

BaTiO 3 single crystals and its interplay with the domain structure 

were investigated by means of electron paramagnetic resonance 

(EPR) spectroscopy and optical microscopy. The examination of 

nanocrystalline ferroelectrics with perovskite structure and the 

determination of their physical and chemical properties are also 

discussed by means of size driven phase tranition from tetragonal-tocubic 

phase. 


EPR 

Ep10 

High-frequency EPR spectroscopy synthetic forsterite crystals 

by doped Cr, Li of ions after irradiation 

Konovalov, A.A. 1 ; Tarasov, V.F. 1 ; Zharikov, E.V. 2 

1 Zavoisky Physical -Technical Institute, Kazan, Russian Federation; 

2 General Physics Institute, Moscow, Russian Federation 

Forsterite (Mg2SiO4) doped by chromium ions is known as an active 

laser medium in the near infrared range, where the laser center is 

the Cr4+ ion in the tetrahedral position. It is known also about laser 

generation of synthetic forsterite co-doped by Cr3+ ions and Li in the 

new for solid-state lasers range of 1,03-1,18 microns [1]. 

As distinct from the Cr4+ ions, which replace silicon in the 

forsterite crystal lattice, the impurity Cr3+ ions replace Mg2+ions in 

octahedral positions named as M1 and M2. In this case, to conserve 

electroneutrality of the crystal compensation of the additional positive 

charge is necessary. In the same magnesium position Cr2+ ions may 

be introduced without charge compensation. This competition prevents 

the formation of centers Cr3+ with a high concentration. 

Increasing of the Cr3+ concentration of and decreasing of the 

Cr2+concentration can be done by co-doping of forsterite by chromium 

and univalent metal ion such as Li. The other method is possibly 

influence on forsterite of high-energy radiation. The method of tunable 

high-frequency EPR spectroscopy was used to investigate the influence 

of high-energy electrons and gamma irradiation of different doses on 

redistribution of the concentration of known chromium ions and the 

formation of new chromium centers. EPR measurements were carried 

out in the frequency range of 64 – 230 GHz at 4.2 К in the Voigt 

geometry as described in [2]. 

For the first time we discovered a new Cr2+ impurity center formed by 

the irradiation of high-energy electron beam or gamma irradiation in 

forsterite co-doped by chromium and thanΔlithium ions. This center 

has a slightly different zero-field splitting the known Cr2+ center but 

has similar angle dependency. We have investigated also dependence 

of relative intensity on doses of gamma irradiation for this new center. 

We found that the relative concentration of the Cr2+ center in the M2 

position is decreased. 

This work is partially supported by the RFBR grant 06-03-16662, 

grant of NIOKR RT 02-2009 and grant of the President of the Russian 

Federation NSh-4531.2008.2. 

1. A.V.Gaister, E.V.Zharikov, V.F.Lebedev, et al. Quantum Electronics, 34 

(8), 693 (2004). 

2. G.S. Shakurov, V.F. Tarasov. Appl. Magn. Reson. 21, 597 (2001.) 

Ep11 

A closed-cycle cryostat for EPR spectroscopy at cryogenic 

temperatures 

Hinderberger, Dariush 1 ; Bauer, Christian 1 ; Bains, Ravi 2 

1 Max Planck Institute for Polymer Research, Mainz, Germany; 

2 Advanced Research Systems , Inc, Macungie, United States 

In this report, we present a closed-cycle cryostat for continuous 

wave (CW) and pulse EPR that is based on continuous compression/ 

expansion of gaseous Helium of high purity. 

The system is easy to use, mechanical vibrations do not interefere with 

pulse EPR measurements and in our setup temperatures of 8.5K can be 

reached (with 4K are possible). Although the mechanical vibrations are 

visible in CW EPR experiments, averaging of several scans removes the 

mechanical noise, as the vibrations are not correlated with the CW EPR 

measurements. 

Our setup drastically reduces the downtime of the spectrometer while 

simultaneously reducing the running cost of an EPR laboratory. 


Ep12 

EPR-Spectroscopic studies of persistent, trivalent and 

mononuclear Pb(III) radicals 

Kurzbach, Dennis 1 ; Hinderberger, Dariush 1 ; Klinkhammer, Karl W. 2 

1 Max Planck Institute for Polymer Research, Mainz, Germany; 2 Johannes 

Gutenberg University, Mainz, Germany 

We present detailed electron paramagnetic resonance (EPR) 

spectroscopic studies of persistent, molecular and mononuclear 

Pb(III) based radicals. The presentation focuses on the homoleptic 

species Pb(Ge(SiMe 3 ) 3 ) 3 with germanium-based ligands as well as 

on heteroleptic radicals substituted by a mixture of silicon-based and 

germanium-based ligands [Pb(Ge(SiMe 3 ) 3 ) 2 Hyp, Pb(Ge(SiMe 3 ) 3 )Hyp 2 (Hyp 

= -Si(SiMe 3 ) 3 )]. Due to enormous contributions of spin-orbit-coupling 

by lead and germanium, these species show larger g-anisotropies than 

the already known, solely silicon-substituted Pb(III) derivates. [1, 2] From 

the reduced (as compared to silicon-only substituted radicals) hyperfine 

couplings a more planar molecular geometry can be deduced with the 

s-p hybrid character of the singly occupied molecular orbital (SOMO) as 

well as contributions of Fermi-contact-interaction decrease. 

Furthermore, the axiality of the respective spectra decreases the more 

germanium-based ligands are replaced by silicon-based ones. This can 

be understood when considering that more spin density is distributed 

to germanium than to silicon. This causes a decrease in symmetry 

of the electronic system in the ligand plane and so leads to higher 

orthorhombicity in the spectra. 

Furthermore, we present a detailed characterization of another new 

species, [PbHyp 2 Ebt (Ebt = -Si(SiMe 3 ) 2 Et)]. This radical distinguishes 

itself through a heteroleptic substitution pattern, which is composed of 

different silicon-based ligands. These ligands differ in size and so lead 

to more pyramidal molecular structures. The respective spectrum shows 

a rhombic g-tensor, indicating that not only the chemical nature of the 

ligands’ atoms but also the geometrical structure of the molecule itself 

determine the spin density distribution in the molecules. 

[1] Klinkhammer et al. Angew. Chem. 2007, 119, 1174 –1177 

[2] Becker et al. Inorganic Chemistry 47, 9965-9978 (2008) 

Ep13 

Analyses of the Heme Ligand–Field Anisotropy in a series of 

Ferric low spin Cytochrome c Mutants 

Kaur, Ravinder 1 ; Bren, Kara L. 1 ; Ensign, Amy A. 1 ; Zoppellaro, Giorgio 2 ; 

Harbitz, Espen 2 ; Andersson, K. Kristoffer 2 

1 University of Rochester, Department of Chemistry, Rochester, United 

States; 2 University of Oslo, Department of Molecular Biosciences, Oslo, 

Norway 

A series of cytochrome c mutants derived from Pseudomonas 

aeruginosa (Pa c–551) and from the ammonia oxidizing bacterium 

Nitrosomonas europaea (Ne c–552) were over–expressed. Point 

mutations were induced in a key residue nearby the Met axial ligand 

(Asn64). The effects were probed by low–temperature electron 

paramagnetic resonance (EPR) and nuclear magnetic resonance (1H 

NMR) spectra analyses. Ne c–552 has a ferric low spin (S=1/2) EPR 

signal characterized by large g anisotropy and gmax resonance at 3.34 

(see figure below), while Pa c–551 (gmax at 3.20) exhibits electronic 

features consistent with very weak axial system. Correlation among 

the electronic fingerprints observed in the over–expressed proteins as 

well as in their mutants (1) shows the occurrence of a linear relation 

ABSTRACTS POSTER


between axial strains (or gmax values) and averaged paramagnetic 

methyl–shifts that stands irrespective of their specific Met dynamics.(2) 

(1) G. Zoppellaro, E. Harbitz, R. Kaur, A. A. Ensign, K. L. Bren and K. K. 

Andersson, J. Am. Chem. Soc., 130, 15348-15360 (2008). 

(2)G. Zoppellaro, K. L. Bren, A. A. Ensign, E. Harbitz, R. Kaur, H.P 

Hersleth, U. Ryde, L. Hederstedt, K. K. Andersson, Biopolymers 

(submitted) 

Ep14 

Avoided level crossing muon spin resonance studies of spin 

probes in soft matter 

McKenzie, Iain 1 ; Dilger, Herbert 2 ; Scheuermann, Robert 3 ; Stoykov, 

Alexey 3 ; Tucker, Ian 4 

1 STFC Rutherford Appleton Laboratory, Chilton, Didcot, United Kingdom; 

2 Universität Stuttgart, Institut für Physikalische Chemie, Stuttgart, 

Germany; 3 Paul Scherrer Institute, Villigen AG, Switzerland; 4 Unilever 

Research and Development, Port Sunlight, United Kingdom 

Avoided level crossing muon spin resonance (ALC-µSR) is a powerful 

magnetic resonance technique for studying free radicals and can 

provide similar information to that obtained with EPR but offers unique 

advantages. Hyperfine coupling constants are determined from the 

position of the resonances in an ALC-µSR spectrum and the resonance 

lineshape can provide information about the motion of the radicals and 

reaction rates. ALC-µSR has been used extensively to study isotope 

effects on the structure and dynamics of radicals, 1 and has recently 

been used as a probe of soft matter systems. 2,3,4 We have used ALC- 

µSR to study the radicals produced by Mu addition to the rod-like 

liquid crystal 4-n-pentyl-4’-cyanobiphenyl (5CB), from which we have 

obtained information about the ordering and molecular dynamics of 

5CB. We have also used ALC-µSR to study the reorientational dynamics 

and local environment of muoniated spin probes formed by Mu addition 

to water-soluble aromatic counterions adsorbed at the oil-water 

interface in lamellar phase dispersions of di-chain cationic surfactants; 

2,3-diheptadecyl ester ethoxypropyl-1,1,1-trimethylammonium chloride 

(DHTAC) and dioctadecyldimethylammonium chloride (DODMAC). The 

results of these experiments will be presented and the advantages and 

disadvantages of the ALC-µSR technique compared with EPR will be 

discussed. 

References 

1. C. J. Rhodes (2002) J. Chem. Soc. Perkin Trans. 2, 1379-1396. 

2. B. W. Lovett et al. (2001) Phys. Rev. B 63, 054204. 

3.vR. Scheuermann et al. (2004) Langmuir 20, 2652-2659. 

4. A. Martyniak et al. (2006) Phys. Chem. Chem. Phys. 8, 4723-4740. 

Ep15 

Characterization of nitrogen-doped carbon nanospheres using 

electron spin resonance 

Keartland, Jonathan; Kunjuzwa, Nikiwe; Coville, Neil 

University of the Witwatersrand, DST/NRF Centre of Excellence in Strong 

Materials, Johannesburg, South Africa 

Carbon nanospheres (CNS) have been synthesized using chemical 

vapor deposition (CVD) methods using a acetylene:argon gas mixture. 

In an attempt to alter both the magnetic properties and the carrier 

concentration, samples were prepared by bubbling the acetylene:argon 

gas mixture through a various nitrogen-rich solutions, with the hope 

that the nitrogen would take up substitutional sites in the carbon 

matrix. Thermogravimetric analysis of the samples have shown that 

appreciable amounts of nitrogen are either adsorbed or incorporated 

in the CNS, and the question arises as to where the nitrogen resides in 

72 

the samples. In an attempt to answer this question we have performed 

ESR measurements at room temperature on the full range of samples 

produced. ESR characterization was performed using a Bruker ESP380E 

X-band spectrometer operating in continuous wave (CW) mode at 

microwave frequencies of approximately 9.75 GHz. Our results show 

that the nitrogen-containing CNS exhibit a strong paramagnetic peak 

at g ≈ 2. This suggests that the nitrogen occupies substitutional sites 

in the carbon network. Moreover, the ESR results have been used to 

determine the optimal nitrogen source, which is a pyridine solution. 

Samples were then prepared using pyridine solutions of varying 

concentrations, and the resulting ESR derivative spectra analysed. 

Estimates of the relaxation times and the g-factor shifts of the nitrogen 

ions were obtained. Relaxation rates increase with increasing nitrogen 

concentration, and small g-factor shifts are observed. Details of the 

sample preparation technique, the ESR results, and complementary 

electrical transport results will be presented. 

Ep16 

Revealing the inner workings of chiral homogeneous catalysts 

using a combined EPR and DFT study 

Van Doorslaer, Sabine 1 ; Vinck, Evi 1 ; Caretti, Ignacio 1 ; Zamani, Sepideh 1 ; 

Murphy, Damien M. 2 ; Fallis, Ian A. 2 

1 University of Antwerp, Physics, Antwerp, Belgium; 2 Cardiff University, 

Chemistry, Cardiff, United Kingdom 

The outcome of an enantioselectively catalyzed reaction is often 

governed by very subtle stereochemical and electronic effects. In order 

to fully understand these effects and thus come to a targeted synthesis 

of novel catalysts, physicochemical characterization methods are 

needed. Here, we show how insight into these catalytic mechanisms 

can be gained from a detailed multi-frequency continuous-wave 

(CW) and pulsed EPR and ENDOR study. We focus on the study of the 

Cu(II) and Co(II) complexes of the Schiff base N,N’-bis(3,5-di-tertbutylsalicylidene)-1,2-cyclohexane-diamine 

(1) introduced by Jacobsen 

and co-workers. Although extensively used in practice, a molecular 

basis for the catalytic working of these catalysts remains uncertain. 

The CW and pulsed EPR analysis of the activation of Co(1) with an 

organic acid reveals the formation of a multitude of species, including 

a Co(III)-bound phenoxyl radical. The interpretation of the EPR data is 

corroborated by DFT computations. Furthermore, binding of different 

enantiomeric ligands to the chiral Co(1) and Cu(1) molecules results in 

subtle but detectable differences in their EPR characteristics that can 

be interpreted in terms of the different stereoselective and electronic 

effects. It will be shown how high-field EPR can play a crucial role in the 

detection of these enantio-specific differences. 

Ep17 

Temperature dependence of Mn 2+ and Fe 3+ ions in Stoichiometric 

LiNbO 3 crystal 

Yeom, Tae Ho; Lee, Soo Hyung 

Cheongju University, Applied Science, Cheongju, Republic of Korea 

LiNbO 3 single crystal is widely used ferroelectric material because of its 

interesting optical properties. Electron paramagnetic resonance spectra 

of Mn 2+ and Fe 3+ paramagnetic impurity ions in stoichiometric LiNbO 3 

single crystal were observed using by EPR spectrometer. Temperature 

dependence of Mn 2+ and Fe 3+ ions was studied in the temperature 

range of 3 K ~ 296 K. Zero field splitting parameter as well as magnetic 

resonance absorption spectra of Mn 2+ and Fe 3+ ions are investigated 

as function of temperature. Temperature dependence of zero field 

splitting parameter of Mn 2+ and Fe 3+ ions was compared with that of 

nuclear quadrupole coupling constant of Li + [1] and Nb 5+ [2] ions in the 

host crystal. Our study supports that that Mn 2+ and Fe 3+ ions in LiNbO 3 


crystal substitute for Nb 5+ ion from the previous reports 

[1] T. H. Yeom, S. H. Choh, and K. S. Hong, J. Korean Phys. Soc. 25, 62 

(1992). 

[2] M. P. Petrov, Sov. Phys. Solid State, 10, 2574 (1969). 

Ep18 

The application of time resolved EPR to the study of energy 

transfer processes in photodynamic therapy 

Iyudin, Vasiliy 1 ; Obynochny, Anatoly 1 ; Salikhov, Kev 1 ; Kruppa, Alexandr 2 

1 Zavoisky Physical -Technical Institute ( ZPhTI ), Kazan, Russian 

Federation; 2 Institute of Chemical Kinetics and Combustion, Novosibirsk, 


In our study we have explored the quenching of the singlet 

oxygen by carotenoids using the time resolved EPR technique. 

We have discovered that the adding of 100 µM β-carotene to the 

21,23-mesotetraphenylporphine - 2,2,6,6,-tetramethylpiperidinyl- 

1-oxyl (TEMPO)-radical sample leads to the unusual character of 

hyperfine interaction (HFI) lines of TEMPO radical. First of all, the sign 

of polarization is positive (absorption) for all three lines, second - all 

lines have different relative intensity (23/48/100%) and third, lines have 

different lifetime. Low field component has a polarization lifetime 2.1 

µs, central component has a polarization lifetime 9 µs and a high field 

component 22 µs relatively to the laser impulse. 

Ander normal conditions, without β -carotene this time dependence 

of HFI lines of TEMPO radical isn’t observed. More than that in 

the degassed sample without β-carotene the emission electron 

spin polarization is observed. Our results could be applied in the 

photodynamic therapy (PDT) for the cancer treatment. 

Ep19 

Dynamics of electron-nuclear polarization transfer at 95 GHz 

detected by ELDOR detected NMR 

Nagarajan, Vijayasarathi; Feintuch, Akiva; Hovav, Yonatan; Vega, Shimon; 

Goldfarb, Daniella 

Weizmann Institute of Science, Chemical Physics, Rehovot, Israel 

The aim of this work is to study the mechanism of Dynamic Nuclear 

Polarization (DNP) at a high field (W-band, 95 GHz, ~3.4 T), specifically 

the initial stage of the depolarization of the electron-spin due to its 

transfer to coupled nuclei. This was measured using the ELDOR– 

detected NMR pulse sequence. In this experiment a long microwave 

pulse with a duration, t HTA , is applied at a frequency ν 1 and after a time 

t d (such that T m < t d < T 1 ) an echo-detection sequence is applied with a 

frequency of ν 2 1 . When ν 1 is on resonance with a forbidden transition 

(ΔM S , ΔM I =±1) the echo intensity decreases. Thus a measurement of 

the echo intensity as a function of Δν=ν 1 -ν 2 gives the NMR spectrum 

of the coupled nuclei. The specific system that was investigated is 

a frozen water/glycerol solution of Gd 3+ (S=7/2) where the intensity 

(negative) of the peak at the 1 H Larmor frequency, Δν=ν 1 -ν 2 =144 

MHz, was measured as a function of the duration of t HTA for different 

microwave powers. The measurements were carried out at 10K in 

the concentration range of 0.5 - 10 mM. We found the following time 

course for high powers (nutation frequency of 8-10 MHz): the 1 H peak 

increases and reaches a maximum within t max ~ 20µs (maximum 

depolarization of the electron-spin allowed transition) and it then 

decays towards equilibrium in a process that involves more than one 

time constant. As the power decreases, the amplitude of the 1 H peak 

decreases, t max increases and the recovery phase is not observed. This 

is a typical behavior expected from the Solid Effect (SE) mechanism. 

This was confirmed by a series of simulations, showing that the shape 

of the recovery phase depends on the electron and nuclear spin 

relaxation times. This behavior was similar throughout the concentration 

range tested confirming the dominance of the solid effect mechanism. 

1. P.Schosseler,; Th. Wacker,; A. Schweiger. Chem.Phys.Lett. 1994, 

224, 319-324. 


Ep20 

Electron spin resonance investigation of the spin ladder system 

(C 5 H 12 N) 2 CuBr 4 

Potocnik, Anton; Zorko, Andrej; El Shawish, Samir; Klanjsek, Martin; 

Arcon, Denis 

Jozef Stefan Institute, Solid State Physics Department, Ljubljana, 

Slovenia 

Lately, the concept of Luttinger liquids (LL) has been widely exploited 

in a variety of one-dimensional (1D) systems, ranging from ultracold 

atoms to carbon nanotubes, quantum wires and antiferromagnets. 

Recently, the first quantitative check of the LL model has been provided 

by the spin-ladder system (C 5 H 12 N) 2 CuBr 4 (BPCB) [1]. In an applied 

magnetic field the spin ladder can be essentially mapped onto a 1D 

system of interacting spinless fermions, with the possibility to control 

LL parameters in a continuous manner with the magnetic field. Any 

eventual magnetic anisotropy can crucially affect the behavior of 

experimental observables in the vicinity of quantum critical points [2]. 

In order to determine the role of magnetic anisotropy in BPCB, we have 

conducted a comprehensive X-band electron spin resonance (ESR) 

investigation on a single-crystalline sample in a wide temperature 

range. Our experimental results show that different line broadening 

mechanisms dominate at different temperatures. The angular 

dependence of the ESR line width and g-factor, compatible with 

previous measurements [3], allowed us to determine the dominant 

anisotropy terms in this compound through theoretical modelling of the 

ESR moments. 

[1] M. Klanjsek et al., Phys. Rev. Lett. 101, 137207 (2008). 

[2] M. Clemancy et al., Phys. Rev. Lett. 97, 167204 (2006); and 

references therein. 

[3] B. R. Patyal et al., Phys. Rev. B 41, 1657 (1990). 

Ep21 

PELDOR on spin labeled ion-channel KcsA 

Endeward, Burkhard 1 ; Prisner, Thomas F. 1 ; Butterwick, Joel A. 2 ; 

MacKinnon, Roderick 2 

1 Goethe University Frankfurt, Frankfurt, Germany; 2 The Rockefeller 

University, New York, United States 

PELDOR (pulsed electron electron double resonance) is a 

magnetic resonance method for distance, orientation, and dynamic 

measurements of two or more paramagnetic centers in macromolecules 

like proteins, RNA, or DNA as well as polymers. We like to demonstrate 

the investigation on an ion-channel KcsA. We studied the mutant KcsA- 

R64C-SL where the spin label is located in a relative large pocked 

outside the membrane. However the PELDOR experiments revealed a 

rigid orientation of the four symmetric spin labels of the tetrameric KcsA 

structure. It seems that there is no structural constrain which might lead 

to that rigidity, but there is no doubt that the only possible explanation 

of the PELDOR time traces is this narrow spin alignment. We did study 

the spin labeled KcsA in detergent as well as in a membrane. The 

preparation in membranes is more demanding than in detergent due 

to the tendency of this membrane protein (like many other proteins) to 

cluster within the membrane. We also demonstrate how to reduce this 

cluster problem. 

ABSTRACTS POSTER


Ep22 

á-Synuclein amyloid fibrils studied by electron paramagnetic 

resonance 

Karyagina, Irina; Becker, Stefan; Riedel, Dietmar; Jovin, Thomas; 

Griesinger, Christian; Bennati, Marina 

Max Plank Institute for Biophysical Chemistry, Göttingen, Germany 

The misfolding of á-synuclein (AS) to a cross β-sheet fibrillar structure 

is associated with pathological conditions in many neurodegenerative 

disorders, including Parkinson’s disease. The mechanism of aggregation 

of AS from its natively disordered state into a well organized fibrillar 

structure involves formation of intermediates, some of which finally 

incorporate into fibrils. In this work, we study the structure of 

α-synuclein in its amyloid fibrillar state by using single and double 

cysteine mutants labelled with MTSL. Broad and weak CW EPR spectra 

were observed with the labelled fibrils. Recently, it was reported that 

this is due to exchange narrowing of the EPR spectrum as a result of 

a strong exchange interaction between the spin probes [1]. A parallel, 

in-register arrangement of monomers in the fibrils was proposed. 

However, the precise structure of α-synuclein fibrils at molecular level 

is still under debate [2, 3]. We prepared a spin-dilution experiment in 

which the double labelled protein was aggregated in different ratios to 

wild type AS. The fibrils were characterized by EM, fluorescence and 

EPR spectroscopy. CW EPR on spin-diluted samples showed features 

indicative of highly immobilised spin labels in the fibrillar state. 4-pulse 

DEER was further used to probe the intermolecular distance and 

distance distribution between the two spin probes in the fibrils. Analysis 

is in progress to determine the orientation of the spin labels in the 

β-sheet and to interpret the distance information in terms of a specific 

molecular structure. 

[1] M. Chen, et al., JBC (2007), 282, p. 24970. 

[2] H. Heise, et al., PNAS (2005), 102, p. 15871. 

[3] M. Vilar, et al., PNAS (2008), 105, p. 8637. 

Ep23 

EPR and magnetization study of ferric green rust- and 

ferrihydrite-coated sands 

Wencka, Magdalena; Jelen, Andreja; Dolinsek, Janez 

J. Stefan Institute, Solid State Physics, Ljubljana, Slovenia 

The high redox reactivity makes the mixed-valence Fe(II)–Fe(III) 

hydroxysalt green rust (GR) compounds important in environmental 

science and technology. GRs play a key role in the corrosion of ironbased 

materials and they can serve as a reductant pool for inorganic 

oxyanions such as nitrates and toxic metals including chromate 

and selenate. These features make GRs promising materials in 

environmental applications, such as reactive filtration processes 

for the elimination and fixation of pollutants like inorganic and 

organic compounds in soils, sediments and contaminated water. 

�� 

reactive iron compound. This ubiquitous iron oxyhydroxide mineral 

is found throughout the soil and water systems and is important to 

many industrial applications. With its high surface area per volume 

and high reactivity, ferrihydrite can interact by surface adsorption 

or by co-precipitation with a number of environmentally important 

chemical species, including arsenic, heavy metals like lead or mercury, 

phosphate, as well as with many organic molecules. Ferrihydrite is thus 

another suitable material for the elimination and fixation of pollutants in 

soils, sediments and contaminated water in reactive filtration processes. 

In the large-scale application such as columns for wastewater 

treatment/filtration, the use or pure bulky iron oxides is relatively 

inconvenient and the coated sand materials are preferable. Coated sand 

particles exhibit an increased active surface area and provide better 

74 

adsorption efficiency of pollutants. We present magnetic properties of 

coated sands by ferric GR of the formula Fe(II)6(1–x)Fe(III)6xO12H2(7– 

3x)CO3 and ferrihydrite minerals, synthesized by different chemical 

methods. Magnetic properties were investigated by EPR spectroscopy 

and by magnetization and magnetic susceptibility measurements. The 

results show that GR-type coatings behave as a superparamagnetic 

system, whereas the ferrihydrite-type coatings are spin-glass-like. 

Ep24 

ESR studies of Eu 0.6 La 1-X Sr X MnO 3 magnetic subsystem separation 

Eremina, Rushana 1 ; Fazlizhanov, Ilshat 1 ; Mukovskii, Yakov 2 

1 Kazan Pisical-Technical Institute, Kazan, Russian Federation; 2 Moscow 

State Institute of Steel and Alloys, Moscow, Russian Federation 

It is known that the magnetoresistance method is highly sensitive. 

In particular, the electron paramagnetic resonance (EPR) method 

allows one to confidently detect the presence of magnetic impurities 

of fractions of percent. In this work, Eu 0.6 La 1- x Sr x MnO 3 polycrystalline 

samples are studied by the EPR method. The nanoscopic magnetic 

structures are found near the phase interface of the ferromagnetic– 

paramagnetic transition on the side of the paraphase. 

ESR measurements were performed in a Bruker CW- spectrometer at 

X- and Q- band frequencies equipped with a continuous N 2 –gas – flow 

cryostat in the temperature region 100 < T < 300 K. The polycrystalline 

samples were placed in quartz tubes. We investigate the ESR spectra in 

Eu 0.6 La 1-x Sr x MnO 3 (x=0.1; 0.13; 0.15; 0.17; 0.2) in the paramagnetic 

regime above magnetic ordering. The spectra consist of a paramagnetic 

signal and ferromagnetic resonance signal at lower resonance fields. 

The line with g eff ~1.97 is associated with the narrowed signal from 

Mn 3+ and Mn 4+ ions and is referred to the paramagnetic region. We 

emphasize that ferromagnetic line is observed in the EPR spectrum of 

Eu 0.6 La 1-x Sr x MnO 3 (x=0.1; 0.13; 0.15; 0.17; 0.2) for temperatures from 

about 260 K to the phase transition temperature. The effective g factors 

of signals at room temperature in low and high magnetic fields are as 

3.0 in the X range, respectively, and 2.4 in the Q range, respectively. 

The magnetic anisotropy of the line position in the X and Q ranges 

does not change. The intensity of ferromagnetic lines is in Q -band 

in 4.5 times smaller than intensity this line in X-band. The existence 

of ferromagnetic regions in the paramagnetic state leads to phase 

separation in manganites. 

Ep25 

DNA conformational flexibility studied by multifrequency 

PELDOR 

Denysenkov, V. 1 ; Marko, A. 1 ; Margraf, D. 1 ; Cekan, P. 2 ; Sigurdsson, S. 

Th. 2 ; Schiemann, O. 3 ; Prisner, T. F. 1 

1 Institute for Physical and Theoretical Chemistry and Center for 

Biomolecular Magnetic Resonance, Goethe-University, Frankfurt am 

Main, Germany; 2 University of Iceland, Science Institute, Reykjavik, 

Iceland; 3 School of Biology, University of St. Andrews, St. Andrews, 


Orientation selective high field pulsed electron double resonance 

(PELDOR) experiments provide in-depth structural information regarding 

distances and relative orientations on radical-pair macromolecular 

complexes in frozen solution as shown previously for example on a 

ribonucleotide reductase dimer [1]. Recently we showed that X-band 

PELDOR experiments with variable probe frequency can also be used 

to determine the mutual orientation between two spin labels that were 

rigidly attached to DNA duplexes [2]. Additionally, we demonstrated 

that a detailed quantitative analysis of such PELDOR time traces allow 

not only to disentangle distance- and angular information, but also to 

explore the conformational flexibility of the macromolecules at their 


freezing temperature [3,4]. 

Here we report that high field PELDOR (6.4 T, 180 GHz) has allowed 

for the first time to prove experimentally the cooperative stretch-twist 

conformational flexibility of double-stranded DNA. Experimental results 

will be shown together with quantitative simulations, and compared with 

previously presented data and models [5,6]. 

1. V. Denysenkov, T.F. Prisner, J. Stubbe, and M. Bennati, Proc. Natl. 

Acad. Sci. USA, 2006, 103, 13386-13390. 

2. O. Schiemann, P. Cekan, D. Margraf, T.F. Prisner, and S. Sigurdsson, 

Angew. Chem. Int. Ed. 2009, 48, 2655-2658. 

3. D. Margraf, B.E. Bode, A. Marko, O. Schiemann, and T.F. Prisner, Mol. 

Phys. 2007, 105, 2153-2160. 

4. A. Marko, D. Margraf, H. Yu, Y. Yu, G. Stock, and T.F. Prisner, J. Chem. 

Phys. 2009, 130, 064 102. 

5. J. Gore, Z. Bryant, M. Nöllmann, M. Le, N. Cozzarelli, and C. 

Bustamante, Nature, 2006, 442, 836-839. 

6. R. Mathew-Fenn, R. Das, and P. Harbury, Science, 2008, 322, 446- 

449. 

Imaging 


Im10 

Image based studies on time course of ADC, signal intensity and 

accuracy on diffusion weighted MR images of cerebral ischemic 

stroke 

Srivastava, Ajai 1 ; Mehrotra, Gopesh 2 ; Bhargava, Satish 2 ; Agarwal, Sunil 3 ; 

Tripathi, Rajendra P 4 ; Tandon, Anupama 2 ; Bhatt, Shuchi 2 

1 University College of Medical Scienecs(University of Delhi)& GTB 

Hospital, Radiology, Delhi, India; 2 UCMS, Radiology, Delhi, India; 3 UCMS, 

Medicine, Delhi, India; 4 INMAS, NMR, Delhi, India 

Time course of ADC and signal intensity changes on DW MR imaging 

in acute ischemic stroke is very dynamic. There is an initial reduction 

in ADC associated with cytotoxic edema, with no change on T2-W 

imaging. At about 6-12 hours, with the onset of vasogenic edema, 

increased T2-weighted signal intensity may become evident. Then, as 

necrosis begins to set in, there is a gradual reversal of ADC change 

because diffusion of water becomes less restricted, and around 3-10 

days ADC pseudonormalises.Twentyfour patients of acute stroke 

underwent diffusion MR Imaging in addition to conventional T1W, T2W 

and FLAIR performed within 12 hours which was repeated at 30 days 

and 90 days in same patients. MR studies were performed on a 1.5 

Tesla MR system. Relative Signal Intensity on DWI and T2W and ADC 

values were obtained. Mean Signal Intensity at b=0 s/mm 2 and on 

diffusion weighted at b=1000 s/mm 2 were significantly higher than 

control values for all time periods. The rSI b=0 significantly increased 

from 1.63 ± 0.20 in the acute stage to 2.19±0.24 in chronic stage 

(p


interval in which the sample becomes saturated with water, the process 

of hardening, and the type of diffusion exhibited. In one case -PVAL/ 

cement- the sample’s behavior is dictated by the competition between 

the leaching of the polymer and the formation of organic acetates. In 

the case of the PVAC/cement sample, the water diffusion trends are 

determined by the evolution of the PVAC hydrolysis reaction on one side 

and by the hardening of the cement on the other. Moreover, we have 

found out that the time for the transition from one type of diffusion to 

another is a function of the temperature at which the hydration takes 

place. 

[1] Y. Ohama (1998) Cem. Concr. Res. 20, 189-212. 

[2] J. Schulze (1999) Cem. Concr. Res. 29, 909-915. 

[3] O. Weichold and M. Möller (2007), Adv. Eng. Mat. 9, 712-715. 

Im12 

Magnetization transfer effect correction in DESIRE based pulse 

sequences 

Reynaud, Olivier 1 ; Webb, Andrew 2 ; Le Bihan, Denis 1 ; Ciobanu, Luisa 1 

1 CEA, NeuroSpin, Paris, France; 2 Leiden University Medical Center, 

Department of Radiology, Leiden, Netherlands 

A potentially mitigating factor in DESIRE based sequences such as 

those described in references [1-3] is that of magnetization transfer. In 

such sequences the presaturation pulses are off-resonance for spins 

located outside the directly saturated region but within the detection 

slice and will lead to a signal increase in the DESIRE images through 

MT effects. In biological tissues the magnetization transfer ratio can 

attain values of 40 to 50% percent which will definitely impede the 

detection of enhancements caused by diffusion or flow. Here we analyze 

the MT effect of the tag pulse on a 2% agar sample and show that 

one can correct for it by adding a magnetization transfer pulse before 

the tag. The idea behind this is to obtain the maximum magnetization 

transfer effect with this MT pre-pulse which will then render the tag 

pulse MT effect free. Our results show that a tag of 900 ms duration 

(60 sinc shaped, 90° pulses, 5 ms duration, 10 ms interpulse delay) 

applied to saturate planes of thicknesses between 200 and 400 µm 

inside a 2 mm imaging slice will produce a 4% MTR on a 2% agar gel. 

This effect can be corrected by applying a train of MT pulses before 

the tag (30 pulses, Gaussian shape, 5 ms duration, 600° flip angle, 1.2 

kHz off water resonance, interpulse separation 100 µs). The optimum 

parameters for the MT prepulse are sample dependent and require 

calibration. Our preliminary in vivo data looking at cerebral flow in rats 

shows tremendous improvement using this strategy. The experiments 

have been performed on a 7T Bruker system. 

References: 

[1] L. Ciobanu, et al. J. Magn. Reson. 170 (2004) 252–256, [2] B. 

Sutton, et al, Magn. Reson. Med., 58:396–401 (2007), [3] M. Weiger et 

al. , J. Magn. Reson. 190 (2008) 95–104. 

Im13 

Functional mapping using variable field proton electron double 

resonance imaging 

Khramtsov, Valery; Shet, Keerthi; Kesselring, Eric; Petryakov, Sergey; 

Sun, Ziki; Zweier, Jay; Samouilov, Alex 

The Ohio State University, Department of Internal Medicine, Columbus, 

OH, United States 

The continuous wave (CW) electron paramagnetic resonance imaging, 

EPRI, is based on spectra measurement at numerous magnetic field 

gradients which requires a long acquisition time. To improve temporal 

resolution we used a proton electron double resonance imaging (PEDRI) 

76 

in combination with a new concept of Variable Field (VF) PEDRI. This 

allows for functional mapping using specific paramagnetic probes (e.g. 

oxygen or pH mapping) within MRI high quality spatial resolution and 

short acquisition time. 

PEDRI with variable field EPR pre-excitation allows for EPR 

spectroscopic information to be obtained along with the distribution 

of the radical within the object. However, complete spectral-spatial 

reconstruction from VF PEDRI requires multiple MRI acquisitions with 

increase of acquisition time by hundred folds. We hypothesized that 

valuable spectral parameters at each pixel can be extracted from a 

limited number of selected PEDRI acquisitions. To prove the concept 

we used the phantom with capillary tubes filled with solutions of 

pH sensitive nitroxide prepared at different pHs. The VF PEDRI were 

acquired at two pre-selected EPR excitation fields which coincide with 

EPR spectral peak positions of protonated and nonpotonated forms of 

the probe. The two images were acquired in 8.8 s providing field of 

view, 30 mmx30 mm, with resolution, 64x64. The ratio of NMR signals 

at each pixel of these two images is pH dependent and was converted 

to pH map with resolution of ~0.1 pH units and a spatial resolution of 

0.5 mm achieved at low NMR field of ~200 G. The data show at least 

10 fold decrease in acquisition time for VF PEDRI compared with EPRI. 

This is particularly important for in vivo applications where stability of 

the paramagnetic probes is limited. Another advantage of VF PEDRI is 

slice selectivity of the functional image which is unavailable in EPRI. 

Note that while concept of functional VF PEDRI was proved using the 

pH probe, it can be applied for studies of other biologically relevant 

parameters of the medium such as redox state, concentrations of 

oxygen or glutathione using specifically designed probes. 

Supported by grants NIH KO1 EB03519 and R01 EB004900. 

Im14 

MRI using an OPENCORE NMR spectrometer 

Inukai, Munehiro; Takeda, Kazuyuki 

Kyoto University, Division of Chemistry, Graduate School of Science, 

Kyoto, Japan 

In this work we demonstrate MRI experiments using an OPENCORE 

NMR spectrometer, which is an open-design, FPGA-based NMR 

spectrometer originally designed for solid-state NMR experiments 

and equipped with three rf channels operational at up to 400 MHz. 

In order to make 1H MRI experiments in static fields of up to 14 T 

feasible, we extended its operational frequency to 600 MHz, and built a 

three-channel field-gradient control extension on the OPENCORE NMR 

spectrometer. 

This work is an example of hardware modification in an opendesign 

NMR spectrometer that has lowered the barrier of building 

spectrometers for those who intend to put their own new ideas in 

practice. 

Im15 

Correlating water and macromolecules across growth rings in 

wood. A multinuclear magnetic resonance imaging study 

Dvinskikh, Sergey 1 ; Henriksson, Marielle 2 ; Berglund, Lars A. 2 ; Furo, 

Istvan 1 

1 Royal Institute of Technology, Department of Chemistry, Stockholm, 

Sweden; 2 Royal Institute of Technology, Department of Fibre and 

Polymer Technology, Stockholm, Sweden 

Wood as material is a composite structure with hierarchical and 

non-random porosity. As has been demonstrated in water-saturated 

greenwood, the total water content anti-correlates with macromolecular 

mass density. In wood dried to equilibrium with ambient humidity the 

situation has been shown to be roughly the opposite: the concentration 


trend of water correlates with the mass density of wood. However, 

previous experiments were not sufficiently accurate and have been 

performed solely by 1 H MRI where, in principle, both water and 

macromolecules can contribute to the same image. 

In this work we investigate the radial distribution of water in wood 

at ambient condition using multinuclear MRI. In contrast to green 

or water-soaked wood, processed construction wood is much less 

suitable for standard MRI because of short relaxation times of “bound” 

water. Hence, solid state MRI methods were applied to assess the 

spatial variation of moisture content in wood. Cubic pieces of ca 6x6x6 

mm 3 size were cut from the wood material (spruce grown in northern 

Sweden) with the edges parallel to the main tree axes. Vacuum dried 

pieces were equilibrated at relative humidities of 95% or 66% over 

respective saturated salt solutions in D 2 O. 

1 H and 2 H NMR spectroscopic and imaging measurements were then 

performed. By comparing the proton and deuterium images we directly 

demonstrate an apparently linear correlation between water and 

macromolecular contents in wood. This is a very direct proof that water 

adsorbed into wood is distributed within and over the cell walls. 

The signal-to-noise ratio we achieved is clearly sufficient for measuring 

variation of various NMR-accessible parameters, such as spin relaxation 

times and water diffusion coefficients, across growth rings. While 

our current data indicate that the amount of adsorbed water per 

macromolecular unit is roughly constant, the molecular mobility of water 

may vary. This issue will be investigated later. 

Im16 

Chemical shift imaging NMR to follow gel formation 

Östlund, Åsa; Bernin, Diana; Nordstierna, Lars; Nydén, Magnus 

Chalmers University of Technology, Department of Chemical and 

Biological Engineering, Göteborg, Sweden 

Polymer gel formation was studied as a function of time by chemical 

shift imaging (CSI) NMR. This technique provides the spatial position of 

each chemical component, and by repeatedly recording sample images 

the chemical rearrangements in the material can be followed in detail. 

The technique follows the same principles as magnetic resonance 

imaging (MRI), but can be accomplished on most of the nowadays 

commercial NMR probes. 

In the study presented here cellulose was dissolved in the well 

characterized solution of tetrabutylammonium fluoride in dimethyl 

sulfoxide. This cellulose solution has the ability to physically crosslink 

into a rigid gel when brought into contact with moisture/water. To study 

the gel formation process the solution was placed in an NMR tube 

and subsequent to the addition of a water-pillar above the solution 

the sample was monitored during 12 hours with the CSI technique. 

By this experimental setup the migration of the compounds, i.e. water, 

TBA+ and DMSO, in the cellulose solution was monitored by collecting 

the 1 H NMR spectrum (i.e. the chemical shift) at each position with a 

resolution of 250 micrometer in the vertical direction of the NMR tube. 

The progression of the gel front was extracted from the CSI profiles as 

a function of time, fitted to the average square displacement of the gel 

front giving the diffusion of 3.4 * 10 -10 m 2 /s. In addition, the release and 

reconstellation of fluoride ions of TBAF were monitored by CSI of the 

19 F NMR signal. It was found that this method is suitable to study the 

release, migration and reassociation of solutes or solvents in materials 

with transient processes on the time scale of 1 hour or longer. 

Metabolomics 


Me10 

NMR Spectroscopy based metabolomic study in mice serum 

under radiation stress 

Moza, Sahil 1 ; Raza Khan, Ahmad 1 ; Rana, Poonam 1 ; Chaturvedi, 

Shubhra 2 ; Khushu, Subash 1 

1 Institute of Nuclear Medicine & Allied Sciences, NMR Research Centre, 

Delhi, India; 2 Institute of Nuclear Medicine & Allied Sciences, Div. of 

Cyclotron and Radiopharmaceutical Sciences, Delhi, India 

Radiation stress, as induced by accident or treatment causes metabolic, 

physiological alterations as well as neuropsychological disorders in 

normal humans and animals. The effect of radiation has to be studied 

for planning of remedial measure in case of event of accident/NBC 

warfare. The NMR spectroscopic-based metabonomic technique has 

been proven to be a powerful tool for characterizing the pathological 

states in animals and humans and can present diagnostic information 

and mechanistic insight into the biochemical effects of the different 

toxins and stressors. Metabolic profiling of serum from irradiated (3 Gy, 

group II and 5 Gy, group III) and control (Group I) mice were investigated 

using NMR spectroscopic-based metabonomic approach. Serum 

samples were analyzed on Bruker Avance, 400.13 MHz Spectrometer 

at 25°C and TSP (1mM) as a reference standard for NMR resonance. 

Fifteen important metabolites were assigned on 1D spectrum which 

included amino acids, organic acids (glycolysis and citric acid cycle 

intermediates), short chain fatty acids, membrane and high energy 

metabolites. In the 1H NMR spectra of serum samples, both the 

irradiated groups exhibited an altered biochemical composition after 5 

days of exposure to radiation. Change in energy metabolism induced by 

radiation was characterised by changes in the levels of lactate, alanine, 

β-OHB, choline, creatine and pyruvate. Although the broad peak regions 

of very low-density lipoprotein (VLDL) and low-density lipoprotein 

(LDL) were suppressed considerably by the CPMG sequence, the peak 

intensities of VLDL and LDL for group II and III were stronger than those 

for the control groups, which indicated alteration in lipid metabolism 

due to radiation stress. Overall, many important metabolic reactions get 

altered during radiation exposure resulting in decreased metabolic rate 

and NMR spectroscopy has given remarkable results. . However, with 

the use of other biochemical, analytical and molecular approach along 

with multivariate analysis, this NMR based metabolomic study will work 

as a platform for developing strategies for minimal invasive radiation 

biodosimetry. 

ABSTRACTS POSTER


78 

Molecular Interactions 

Mi10 

Structure and function of the OCRE domain involved in 

alternative splicing regulation 

Mourão, André 1 ; Bonnal, Sophie 2 ; Miñana, Bela 2 ; Valcárcel, Juan 3 ; 

Sattler, Michael 4 

1 Helmholtz Zentrum München, Technische Universität München, EMBL 

Heidelberg, München, Germany; 2 Centre de Regulacio Genomica and 

Universitat Pompeu Fabra, Barcelona, Spain; 3 Centre de Regulacio 

Genomica and Universitat Pompeu Fabra, ICREA, Barcelona, Spain; 

4 Helmholtz Zentrum München, Technische Universität München, 

München, Germany 

RBM5 is a putative tumor suppressor gene frequently inactivated 

in cancers of the lung and other tissues and down-regulated upon 

oncogenic Ras activation. The gene is overexpressed in breast 

tumors or upon expression of the HER-2 oncogene in breast and 

ovarian cancer cell lines. Its main protein product has been detected 

in pre-spliceosomal complexes and modulates cell proliferation and 

Fas-mediated apoptosis. RBM5 was identified as being a component 

of complexes involved in 3’ splice site recognition and as a regulator 

of Fas exon 6 alternative splicing, switching between pro- and antiapoptotic 

forms of the receptor. Contrary to classical mechanisms of 

splicing regulation, RBM5 does not affect early events of splice site 

recognition that lead to exon definition. Instead, RBM5 inhibits the 

transition between pre-spliceosomal complexes assembled around exon 

6 to mature spliceosome assembly on the flanking introns. 

We have found that an OCRE (OCtamer REpeat of aromatic residues) 

domain is important for RBM5 function in vivo and contacts components 

of the U4/5/6 tri-snRNP. Using NMR spectroscopy, we determined the 

three-dimensional structure of this domain in solution. The RBM5 OCRE 

domain represents a novel â-sheet fold. Specific aromatic residues of 

this domain forming a hydrophobic pocket are key determinants both for 

the interaction with protein partners and for splicing regulation in vivo 

. Replacement of specific tyrosine residues by other aromatic residues 

(phenylalanine or tryptophane) is tolerated and does not disrupt either 

the interaction with protein partners or the function of the protein in vivo 

. In contrast, substitution to alanine or threonine leads to local unfolding 

and disrupts the function of the domain. These structural and mutational 

analysis identifies a binding site for putative ligands and suggests 

that aromatic residues on the surface of the OCRE domain mediate 

molecular interactions required for splicing regulation. 

Mi11 

Identification of a hexapeptide with strong antiviral activity 

against the SARS coronavirus and characterization of its 

binding mode by saturation transfer difference (STD) NMR 

spectroscopy 

Struck, Anna-Winona 1 ; Pfefferle, Susanne 2 ; Axmann, Marco 1 ; Drosten, 

Christian 3 ; Meyer, Bernd 1 

1 University of Hamburg, Faculty of Sciences, Department of Organic 

Chemistry, Hamburg, Germany; 2 Bernhard Nocht-Institute for Tropical 

Medicine, Clinical Virology, Hamburg, Germany; 3 University Hospital 

Bonn, Faculty of Medicine, Institute of Virology, Bonn, Germany 

A defined receptor binding domain (RBD) on the viral spike protein (S) 

mediates the attachment of SARS Coronavirus to its cellular receptor, 

angiotensin converting enzyme 2 (ACE2). We have analyzed binding of 

the SARS S-protein with ACE2 by SPR and STD NMR. From a peptide 

library a hexapeptide from Tyr438 to Leu443, Tyr-Lys-Tyr-Arg-Tyr-Leu 

(YKYRYL), of S-protein was identified to have binding affinity to ACE2 

(K D = 46 µM). This peptide has also strong antiviral activity and can 

suppress viral proliferation completely at a concentration of 10mM. 

STD NMR spectroscopy was used to detect the interaction of YKYRYL 

and related peptides with the receptor protein ACE2. Close contacts of 

the aromatic tyrosine residues to the receptor were identified. Also, Arg 

441 interacts strongly with the S-protein. 

Mi12 

DNA condensation induced by polyamines as observed by 

electron paramagnetic resonance spectroscopy 

Hinderberger, Dariush; Kizilsavas, Gönül; Kurzbach, Dennis 


Two approaches are investigated for gene therapy, viral and non-viral 

delivery vectors. The disadvantage of viral vectors is their ability to 

cause harmful mutations of the genome or even cancer when applied 

to the patient. In this regard, non-viral vectors are less dangerous, but 

so far also less efficient. The challenge is thus to optimize non-viral 

DNA transferring systems. One potential non-viral delivery system can 

be found in nature, in sperm cells and bacteriophage-heads. The DNA 

packing in sperm cells is induced by protamines which replace histones 

in the haploid phase of spermatogenesis and are responsible for DNA 

condensation and stabilization. 

Our work focuses on the protamine spermine and its dendritic 

derivatives. To optimize non-viral DNA transferring systems, a deep 

understanding of the interactions on the molecular scale and the 

dynamics of the condensation is required. To this end, magnetic 

resonance methods such as nuclear magnetic resonance (NMR) or 

electron paramagnetic resonance (EPR) can give valuable insights. EPR 

spectroscopy detects unpaired electron spins in radicals that are either 

intrinsic (e.g. paramagnetic metal centers in proteins) or need to be 

added synthetically as so-called spin-labels. Our results show that upon 

spin-labeling of the spermine-derived condensing agents with nitroxide 

radicals, EPR-spectroscopy delivers insight into the interaction (mainly 

of ionic nature) of DNA and the oppositely charged spermine-based 

molecules. Continuous-wave EPR and methods of pulse EPR (HYSCORE 

and DEER) were used to characterize the dynamics of the interactions in 

solution leading to the polyamine-induced condensation of DNA and the 

subsequently formed structures. 

Mi13 

Direct NMR detection of hydrogen bonds and quantification 

of h J-coupling constants in an acylguanidine bisphosphonate 

complex 

Drettwan, Diana; Federwisch, Guido; Kleinmaier, Roland; Gschwind, 

Ruth M. 

Universität Regensburg, Organic Chemistry, Regensburg, Germany 

The H-bonding network of arginine based on specific interactions with 

the corresponding receptor moieties is crucial for many biological and 

pharmaceutical interactions, because of the various possibilities of 

the guanidines to build charge assisted H-bond networks. The effect 

of acylation of guanidinium moieties on the binding mode and H-bond 

strengths was investigated, because acylguanidines provide significantly 

improved pharmacokinetics, activities and selectivities in several 

receptor families [1]. However, the directed modification of such H-bond 

networks is still very difficult due to a missing experimental accessibility 

of H-bonds. 

Using an artificial arginine receptor, initially developed by T. Schrader 

[2], we were able to detect directly H-bonds to individual acylguanidine 

protons with the help of 1D- and 2D- 1 H, 31 P-HMBC [3]. The direct NMR 

detection and quantification of 1D, 2D and 3D correlations caused by 

2h JHP and, for the first time in non-biomolecules, 3h J NP -couplings [3], 


leads ultimately to an insight into the spatial arrangement of the NH-OP 

H-bonds and indicates an end-on binding mode. 

[1] P. Ghorai, A. Kraus, C. Gotte, P. Igel, E. Schneider, D. Schnell, G. 

Bernhardt, S. Dove, M. Zabel, S. Elz, R. Seifert, A. Buschauer, (2008) J. 

Med. Chem. 51, 7193 

[2] T. Schrader, (1997) Chem. Eur. J. 3, 1537 

[3] R. M. Gschwind, G. Federwisch, R. Kleinmaier, D. Drettwan , (2008) 

J. Am. Chem. Soc. 130, 16846 

Mi14 

NMR of amorphous materials - from local interactions to the 

structure of materials 

Shenderovich, Ilja G. 

Freie Universitaet Berlin, Berlin, Germany 

Whether X-ray or NMR suits more to elucidate the structure of crystals 

is a debatable question. For amorphous materials NMR is out of 

competition. Here we describe a strategy employed to study amorphous 

materials using NMR spectroscopy. In all cases the starting point is 

chemically active sites of the studying materials. To be more precise 

they are the sites able to form hydrogen bonds either between each 

other or with guests. In some cases the inspection of the properties 

of these active sites makes it possible to describe the structure of the 

materials. The first example is investigation of morphology and potential 

chemical reactivity of highly ordered porous silica.[1] For the most 

ordered of them NMR provides a checklist for atomic-scale models of 

these amorphous but periodically structured porous materials.[2] These 

details are not only of academic interest but also of importance for 

applications of these materials as catalysts and host materials. Another 

example is a paper made of aminocarboxyl group grafted cellulose. It 

is shown that the functional aminocarboxyl groups form zwitterionic 

dimers structured in a dimer ribbon or tetramers. This zwitterionic state 

is responsible for a wet-tensile-strength improvement of the paper 

made of the grafted cellulose. 

[1] I.G. Shenderovich, G. Buntkowsky, A. Schreiber, E. Gedat, S. Sharif, J. 

Albrecht, N.S. Golubev, G.H. Findenegg, H.-H. Limbach J. Phys. Chem. B 

2003, 107, 11924. 

[2] I.G. Shenderovich, D. Mauder, D. Akcakayiran, G. Buntkowsky, H.-H. 

Limbach, G.H. Findenegg J. Phys. Chem. B 2007, 111, 12088. 

[3] R. Manriquez, F.A. Lopez-Dellamary, J. Frydel, T. Emmler, H. Breitzke, 

G. Buntkowsky, H.-H. Limbach, I.G. Shenderovich J. Phys. Chem. B 

2009, 113, 934. 

This study is supported by the Russian Ministry of Education 

and Science (RNP 2.1.1. 485), the Russian Foundation of Basic 

Research (Project 09-03-91336-NNIO_a), and the Deutsche 

Forschungsgemeinschaft (Sfb 448). 

Mi15 

Fragment based screening of human blood group B 

galactosyltranferase (GTB) using NMR and SPR 

Landström, Jens 1 ; Rademacher, Christoph 2 ; Sindhuwinata, Nora 2 ; 

Widmalm, Göran 1 ; Peters, Thomas 2 

1 Stockholm University, Organic Chemistry, Stockholm, Sweden; 

2 University of Luebeck, Institute for Chemistry, Luebeck, Germany 

Glycosyltransferases (GTs) are an important enzyme family which is 

involved in the biosynthesis of oligosaccharides, polysaccharides and 

glycoconjugates. The human blood group B galactosyltransferase (GTB) 

catalyzes the transfer of galactose from uridine diphosphate galactose 

(UDP-Gal) to the O3 position of the terminal galactose residue of 

H-antigen acceptors to finalize the formation of the human blood group 

B-antigen. 

Finding inhibitors for this process could be of importance for directed 

metabolic engineering with a perspective to be of help in blood 

transfusions and organ transplantations. 

We are presenting results from a fragment based screening of GTB 

against approximately 500 small molecule fragments from the RO5 

Maybridge library. A combination of STD NMR, T1ñ filtered NMR and 

Surface Plasmon Resonance (SPR) was used to study the interactions 

between GTB and the small molecules. Based on these data it is now 

possible to design novel high affinity inhibitors for the GTB. Moreover, 

activity assays have also been employed to study the fragments’ 

specificity and inhibitory effect on glycosylation. 


Mi16 

NMR study of polymer-solvent interactions during phase 

separation in solutions of thermoresponsive polymers 

Spevacek, Jiri 1 ; Hanykova, Lenka 2 ; Starovoytova, Larisa 1 ; Kourilova, 

Hana 2 ; Labuta, Jan 2 

1 Institute of Macromolecular Chemistry AS CR, v.v.i., Prague, Czech 

Republic; 2 Faculty of Mathematics and Physics, Charles University, 

Prague, Czech Republic 

Some polymers, including poly(vinyl methyl ether) (PVME) and poly(Nisopropylmethacrylamide) 

(PIPMAm), exhibit in aqueous solutions a 

lower critical solution temperature (LCST), i.e., they are soluble at 

lower temperatures, but their heating above the LCST results in phase 

separation. On the molecular level, phase separation is a macroscopic 

manifestation of a coil-globule transition followed by aggregation and 

formation of so called mesoglobules. Here we report some results on 

hydration and other polymer-solvent interactions as obtained with D2O 

solutions of PVME and PIPMAm using NMR spectroscopy. 

For PVME solutions (c = 2-10 wt%), 1H NMR relaxation revealed that a 

certain portion of water is bound in mesoglobules [1]. NMR relaxation 

and diffusion measurements have shown that bound water exists also 

in mesoglobules of PIPMAm. In this case two types of bound water 

molecules were detected in relaxation experiments with diffusion filter 

and assigned to water molecules bound inside mesoglobules and on 

their surface. A fast exchange between free and bound water molecules 

(residence time ~1 ms) was observed both for PVME and PIPMAm 

solutions. In contrast, for highly concentrated PVME/D2O solutions 

(c ≥ 20 wt%) a slow exchange (residence time 2.1 s) was found. A 

three orders of magnitude slower exchange in highly concentrated 

PVME solutions is in accord with the optical microscopy findings that 

mesoglobules are here approx. 30 times larger than in PVME solutions 

with c ≤ 10 wt%. 

We also used NMR spectroscopy to investigate PIPMAm solutions in 

D2O/ethanol (EtOH) mixtures. The phase separation was observed 

even at 298 K for certain compositions (~40 vol% of EtOH) of the 

mixed solvent as consequence of the cononsolvency effect. Presence 

and absence of bound EtOH in mesoglobules induced by temperature 

and cononsolvency, respectively, was shown by 13C NMR relaxation 

measurements. 

Acknowledgment: This work was supported by the Grant Agency of CR 

(projects 202/09/1281 and 203/07/P378), Grant Agency of the Charles 

University (project 27508/2008) and AS CR (project 40500505). 

[1] J. Spevacek, L. Hanykova, L. Starovoytova, Macromolecules 

2004;37;7710. 

ABSTRACTS POSTER


Mi17 

New perspectives for STD NMR and transfer NOE spectroscopy 

for the characterization of dispersant-nanoparticle interactions 

in colloidal dispersions 

Szczygiel, Agnieszka 1 ; Timmermans, Leo 2 ; Fritzinger, Bernd 1 ; Martins, 

Jose C. 1 

1 Universiteit Gent, Dept. Organic Chemistry / NMRSTR, Gent, Belgium; 

2 Agfa Gevaert, Mortsel, Belgium 

Saturation Transfer Difference (STD) NMR and transfer NOE 

spectroscopy are well established techniques in biomolecular NMR 

spectroscopy as a powerful tools to screen for interactions between 

small molecule ligands and their macromolecular targets. 1 Despite 

their considerable success, few - if any - reports applying STD NMR 

have appeared outside the biomolecular NMR realm. HEre, we first 

demonstrate new application perspectives for STD NMR to study 

the organization of dispersant molecules at nano-crystalline organic 

pigment surfaces in colloidal dispersions, typically employed in inks and 

paints. 

The interaction between SDS molecules and quinacridone pigment 

particles (Pigment Red 122) in D 2 O is used as a simplified model 

system. The organic pigment provides a dense network of tightly 

coupled proton spins that can be selectively irradiated. The saturation 

is transferred to any ligand that adsorbs to the surface under fast 

exchange conditions. Using the concentration dependence of the STD 

amplification factors, a surface rearrangement of the surfactant from 

flat-on to hemi-cilindrical structures is demonstrated. Competition 

experiments with tetramethylammonium and the use of protonated/ 

deuterated SDS mixtures confirm this picture. General applicability is 

demonstrated in acetone, by monitoring the interaction between a low 

molecular weight polymer and PR122 particles . 

Transfer NOESY is proposed for the case where the nanoparticle does 

not feature a proton network, for instance for colloidal dispersions 

of semiconductor nanocrystals. We show 2 that useful information is 

obtained when fast exchange of the ligand to the surface prevails and 

other parameters fail to report on the interaction because their averaged 

value is dominated by the free ligand. 

While there are clear differences with biomacromolecules, we believe 

that STD NMR and transfer NOESY show considerable promise in the 

study of dispersant – nanoparticle interactions, providing information at 

the molecular level which is hard to come by using other spectroscopic 

techniques. 

1 Meyer, B., Peters, T., Angew. Chem. Int. 2003, 42, 864-890. 

2 Martins, J.C. et al, J. Am. Chem. Soc. 2009, 131, 3024-3032. 

Mi18 

Structure, dynamics and ligand interactions that describe an 

allergen 

Rundqvist, Louise 1 ; Tengel, Tobias 1 ; Alcocer, Marcos 2 ; Eva, Selstam 3 ; 

Schleucher, Jürgen 1 ; Larsson, Göran 1 

1 Umeå University, Medical Biochemistry and Biophysics, Umeå, 

Sweden; 2 University of Nottingham, Division of Nutritional Sciences, 

Loughborough, United Kingdom; 3 Umeå University, Umeå Plant Science 

Centre, Umeå, Sweden 

Ber e 1 is the major allergen present in brazil nuts. It belongs to the 

2S albumin family, a class of proteins frequently represented in food 

allergy. Here we present the three-dimensional structure determined 

by solution state NMR. We have also measured 15 N relaxation to assess 

the dynamic properties of Ber e 1 in solution. However, the purified 

protein alone does not describe the molecular origins of the allergenic 

properties; the presence of endogenous lipids is required for an allergic 

reaction, which has been shown in mouse models. The active lipid 

80 

fraction has been analysed by solution state NMR using the PH-COSY 

experiment, revealing a mixture of different phospholipids. Ber e 1 

�� 

complex suitable for solution state studies of both structure and 

dynamics. In addition, we show that Ber e 1 is able to bind copper 

with high affinity to a specific binding site. The various ligand binding 

properties of Ber e 1 is compared to SFA8, a homologous protein 

originating from sunflower seeds which far less allergenic, yet very 

similar in three-dimensional structure. By defining the differences 

between the free and lipid bound state of Ber e 1, as well as differences 

between a strong and weak allergen, we may be able to describe the 

requirements for a protein to become allergenic in general. 

Mi19 

Ligand-Altered dynamics in allosteric gene regulation by a 

macromolecular protein ring 

Kleckner, Ian 1 ; McElroy, Craig 1 ; Sachleben, Joseph 1 ; Xu, Jiageng 1 ; 

Gollnick, Paul 2 ; Foster, Mark 1 

1 Ohio State University, Columbus, Ohio, United States; 2 University at 

Buffalo, Buffalo, NY, United States 

The biosynthesis of tryptophan is regulated in Bacilli by a myriad 

of macromolecular interactions that respond to cellular metabolite 

concentrations. The 91 kDa ring-shaped undecameric (11-mer) trp 

RNA-binding attenuation protein (TRAP) senses the levels of free 

tryptophan and allosterically regulates gene expression through 

interactions with nascently transcribed mRNA. In addition, the RNA 

binding activity of ligand-activated TRAP can be inhibited via its 

interactions with the multimeric anti-TRAP protein (AT). Backbone amide 

and sidechain methyl relaxation measurements of ps-ns and µs-ms 

timescale motions provide detailed insights into the structural and 

dynamic basis for allosteric control of gene expression in this complex 

and tightly regulated biochemical system. 

NMR studies of ILV-methyl labeled TRAP have allowed site-specific 

mapping of motional modes and microstates accessible to the apo 

protein, and show that most of these modes disappear upon ligand 

binding. By fitting methyl relaxation dispersion data recorded at multiple 

fields and temperatures to a two-state model, we have identified groups 

of residues with synchronized exchange rates, as well as residues 

exhibiting uncorrelated dynamics. Further, we find that the “invisible” 

conformations sampled by apo-TRAP resemble neither the apo state 

nor the ligand bound state. 

In addition, we have determined the solution structure and dynamic 

behavior of the TRAP inhibitor, AT, in its dodecameric and trimeric 

states. Chemical shift perturbations coupled with other biophysical 

studies provide insights into the mechanism by which AT prevents RNA 

binding by Trp-activated TRAP. 

These findings have implications for the mechanism of ligand binding 

and the kinetics and thermodynamics of the regulatory switch. 

Mi20 

In situ muSR and NMR investigations of methanol dissociation 

on carbon supported nanoscaled catalyst 

Suleimanov, Nail 1 ; Khantimerov, Sergei 1 ; Kukovitsky, Eugene 1 ; 

Scheuermann, Robert 2 ; Herlach, Dierk 2 ; Gnezdilov, Oleg 1 ; Locshin, 

Alexander 1 

1 Zavoisky Physical-Technical Institute of Russian Academy of Sciences, 

Kazan, Russian Federation; 2 Paul Scherrer Institute, Laboratory for 

Muon Spin Spectroscopy, Villigen PSI, Switzerland 

In direct methanol fuel cell the methanol-to-hydrogen decomposition 

goes through the formation of intermediate reaction products, such 

as methoxide, formaldehyde, carbon monoxide and hydrogen. In this 


eport in situ muSR and NMR experiments on the investigation of 

methanol decomposion on carbon supported nanoscaled catalyst are 

presented. The short introduction on the application of muSR and NMR 

methods to study the structure and dynamics of molecules adsorbed 

on the surface of catalytic and porous materials is given. In initial Pt- 

Ru/ Vulcan XC-72 carbon black the diamagnetic muon signal is only 

observed by transverse field muSR. It is found that the asymmetry of 

diamagnetic signal tends to increase with increase in temperature. 

Then the methanol was deposited at Pt-Ru/Vulcan XC-72 in the amount 

corresponding to the monolayer coverage and measurements were 

done at a such samples. It is found that the asymmetry of diamagnetic 

signal is decreased in comparing with the Pt-Ru/ Vulcan XC-72. The 

results obtained indicate that there is the fast relaxing muons in Pt-Ru/ 

Vulcan XC-72 /MeOH. Avoided level crossing muon spin resonance 

measurements reveal the resonance line at magnetic field 2.018 T, we 

assign to transient CH 2 �� 

obtained allow us to estimate in situ the contribution of chemical 

decomposition reaction process for the formaldehyde - carbon oxide 

- hydrogen conversion on Pt-Ru/Vulcan XC-72 catalytic material. NMR 

spectrum of the methanol covered Vulcan XC-72/Pt-Ru consists of 

three broad resonance lines assosiated with the methanol molecule 

and chemisorbed hydrogen. The interesting feature is the fact that NMR 

spectra of electrochemically hydrogenated carbon nanotubes exhibit the 

similar resonance line as observed at methanol decomposition on Pt- 

Ru/Vulcan XC-72. The dynamics of different groups of protons and rate 

constants of intermediate products is discussed. 

Acknowledgement 

The support of the Ministry of Education and Science of the Russian 

Federation through the project adtp 2.1.1/4982 and Russian Foundation 

for Basic Research, grant 07-08-12196 is gratefully acknowledged 

Mi21 

The application of NMR techniques to study the reactivity of 

biomolecules and drugs in nanosized aggregates 

Leshina, Tatyana; Kruppa, Alexandr; Polyakov, Nicolai; Petrova, Svetlana; 

Kornievskaya, Valeria; Schlottgayer, Anna 

Institute of Chemical Kinetics and Combustion SORAN, Laboratoty of 

Magnetic Phenomena, Novosibirsk, Russian Federation 

In recent years nanosized organic host-guest complex and micelles 

has become an intensively pursued field of organic and medicinal 

chemistry. Highly interesting are molecular complex formation involving 

macrocyclic, biologically active compounds isolated from biological raw 

materials. Such agents can protect parent substances from metabolic 

decay and provide for their prolonged action. But the influence of 

complexation is not restricted to only the protection role. Changes of 

some physicochemical characteristics of biologically active molecules 

due to complexation can result in changes of their therapeutic activity. 

One of the main questions in the investigation of chemical processes 

in the organized media (nanosized supramolecular complexes, 

micelles, liquid crystals, e.a.) is the elucidation of mechanisms of 

these aggregates influence on the reactivity of the “guest” molecules. 

To answer this question we present a new approach based on 

combination of NMR and spin chemistry methods. We have found the 

correlation between NMR characteristics of complexes and micelle 

in solution at different concentrations of “host” molecules and the 

efficiency of separate stages of radical reactions of biologically relevant 

compounds in complexes and micelles. The control of complexes and 

micelles formation was made by independent method (NMR relaxation 

measurement). Radical reactions efficiency was measured by CIDNP 

techniques, EPR and UV spectroscopy. 

This approach is illustrated by examples of the demonstration of 

the influence of complexes and micelles of natural amphiphilic 

compound, glycyrrhizic acid (GA), complexes of beta-cyclodextrin 

and arabinogalactan on carotenoids antioxidant properties . It was 

also shown the variance of the reactivity of aminoacids and several 

cardiovascular drugs in model redox proccesses in GA and cyclodextrin. 

It is suggested the abovementioned alternations result in the influence 

of supramolecular environment on molecular dynamics of radical pairs. 

For example, the ratio of contributions of CIDNP from radical – ion pairs 

and free radical pairs in complexes differ from that one in solution. 

The work is supported by RFBR grant 08-03-00372. 


Mi22 

Ligand binding and counterion condensation investigated by 

electrophoresis NMR 

Böhme, Ute; Scheler, Ulrich 

Leibniz Institute of Polymer Research Dresden, Dresden, Germany 

The effective charge of a macromolecule is often significantly smaller 

than the nominal charge, because of condensed counterions. Usually 

the energy of the counterions is insufficient to escape the electric 

field of the macromolecule, therefore a fraction of the counterions 

condenses, lowering the effective charge. It is seen in both diffusion 

and electrophoresis NMR [2]. The effective charge is determined in a 

combination of diffusion and electrophoresis NMR [3-5] The effective 

charge is smaller, than the charge determined from titration, because 

during titration the condensed counterions are successively replaced as 

seen in BSA [5]. As simple models for globular molecules dendrimers 

with charge groups on the surface or in bulk have been investigated, 

showing counterion condensation even at low molecular weights. 

As an example for ligand binding glutamic acid and small peptides 

interacting with PDADMAC, a strong polyelectrolyte, have been studied. 

The charge of PDADMAC is independent of pH, while that of glutamic 

acid and the peptides increases by magnitude with pH. At high pH both 

acid groups are dissociated and an effective charge of –2 is found. 

When the charge of glutamic acid increases, an increasing fraction 

binds to the polyelectrolyte, despite the fact, that the acid becomes 

more hydrophilic at the same time. That proofs, that glutamic acid binds 

to the polycation only via electrostatic interaction. 

[1] G. S. Manning, 1974. 

[2] U. Böhme, U. Scheler, Macro. Symp., 211, (2004) 

[3] U. Böhme, U. Scheler, Col.Surf.A, 222, (2003) 

[4] U. Scheler, in ‘Handbook of Polyelectrolytes and their applications’, 

American Scientific Publishers (2002), Vol. 2, 173 

[5] K. Grass, U. Böhme, U. Scheler, H. Cottet, C. Holm, PRL 100, (2008) 

[6] U. Böhme, U. Scheler, CPL 435, (2007) 

Mi23 

Structure and ligand interactions of two Tudor domains 

Friberg, Anders 1 ; Oddone, Anna 2 ; Corsini, Lorenzo 2 ; Mourao, Andre 2 ; 

Sattler, Michael 1 

1 Biomolecular NMR, Technische Universität München, Garching, 

Germany; 2 EMBL, Heidelberg, Germany 

The Tudor domain is a small beta-sheet fold domain that binds ligands 

containing methylated lysines and arginines. Here, we present the NMR 

structure of the Tudor domain of Polycomb-like (Pcl) as well as crystal 

structure of Tudor-SN, both from Drosophila melanogaster. 

The Pcl protein is important for epigenetic regulation of certain 

developmental genes, i.e. HOX genes. Its presence is needed in the 

cell to reach high levels of trimethylation of histone tails (H3-K27), 

which at a later stage leads to condensed chromatin and silencing 

of genes in that particular region. We investigated binding of various 

potential ligands of Pcl Tudor by NMR, however, we could not detect 

any interaction with methylated arginine or lysine ligands. The threedimensional 

structure of the Pcl Tudor domain provides a rationale for 

ABSTRACTS POSTER


these results, as it lacks an intact aromatic cage. A hydrophobic patch 

on the surface of the Tudor domain might serve as an interaction site for 

other domains of Pcl or other proteins. 

The Tudor-SN protein has been implicated in a variety of cellular 

processes, like transcription initiation, processing of edited dsRNA, 

and more recently it has been linked to splicing regulation. We have 

solved the crystal structure of an extended Tudor domain of Tudor-SN 

from Drosophila melanogaster at a resolution of 2.1 Å. NMR secondary 

chemical shifts, relaxation data and residual dipolar couplings indicate 

that the solution and crystal structures are similar. Binding sites and 

affinities were characterized by chemical shift perturbations (CSPs). 

We show that the aromatic cage of the Tudor domain of Tudor-SN 

specifically binds a peptide containing symmetrically dimethylated 

arginines (sDMA), while the same peptide comprising non-methylated 

arginines does not show CSPs. Recognition of sDMA in the C-terminal 

tails of spliceosomal Sm proteins suggests how Tudor-SN may interact 

with small nuclear ribonucleoprotein-particles (snRNPs) during the 

regulation of splicing. 

Mi24 

Study of the interaction of La(III) complexes of DOTA/DTPA- 

Glycoconjugates with RCA120 using the high resolution 

saturation transfer difference (STD) NMR technique 

Geraldes, Carlos F.G.C 1 ; Teixeira, João C. 1 ; Martins, José A. 2 ; André, 

João P. 2 ; Cañana, J. 3 ; Jiménez-Barbero, Jesus 3 

1 University of Coimbra, Department of Biochemistry, Coimbra, Portugal; 

2 University of Minho, Chemistry Center, Braga, Portugal; 3 CIB-CSIC, 

Complutense University, Department of Protein Science, Madrid, Spain 

The study of ligand-receptor interactions using high resolution NMR 

techniques became much easier and with wider applications with the 

development of the Saturation Transfer Difference (STD) technique 

[1]. With such a protocol it is possible not only to rapidly determine 

if there is an interaction between a relatively small compound and a 

protein but also to screen out the ligand interaction specificity at atomic 

level, a characterization known as group epitope mapping (GEM). Our 

aim in this work is to prove the interaction between Gd(III) -based 

glycoconjugate complexes, to be used as potential MRI contrast agents, 

with a soluble target protein and to characterize the ligand-protein 

binding interactions at an atomic level. We screened the interactions 

between the La(III) complexes of a group of DOTA-glyconjugates, 

namely DOTAGal, DOTALac, DOTA(Lac)2, with the 120kDa lectin 

Ricinus communis agglutinin (RCA120), used as a model for the 

asyaloglycoprotein receptor (ASGP-R)[2]. 1H NMR STD titrations allowed 

the association constants of the chelates to the protein to be obtained. 

The % STD values obtained for the different chelate protons were 

used together with the molecular docking program AUTODOCK 4 [3] to 

model the details of the chelate-protein interactions at the atomic level. 

Our results were consistent with the expectations based on previous 

literature [2]. They prove that LnDOTA-glyconjugates efficiently bind 

RCA120 mainly through the galactosyl residue. Some interactions occur 

between the hydrophobic linker arms of the chelates and hydrophobic 

patches at the surface of RCA120. 

References 

[1] Mayer, M.; Meyer, B., J. Am. Chem. Soc., 2001, 123, 6108-6117. 

[2] André, J. P.; Martins, J.A.; et al., Chem. Eur. J., 2004, 10, 5804- 

5816. 

[3] Goodsell, D. S.; Morris, G. M.; Olson, A. J. Journal of Molecular 

Recognition. 1996, 9, 1-5 

Acknowledgments 

We thank the support from F.C.T. Portugal (project PTDC/ 

QUI/70063/2006), COST D38 and the EU-Project EMIL. 

82 

Mi25 

Probing the hepatocyte growth factor/scatter factor - 

Glycosaminoglycan interaction by NMR 

Johansson, Conny M; Uhrín, Dušan 

University of Edinburgh, School of Chemistry, Edinburgh, United 

Kingdom 

Hepatocyte growth factor/scatter factor (HGF/SF) is a key hormone in 

many physiological processes such as wound healing and cancer. HGF/ 

SF exerts its functions through binding the tyrosine kinase receptor 

c-Met. To be able to effectively bind and activate c-Met, HGF/SF 

interacts with a cell surface glycosaminoglycan (GAG) such as heparan 

sulfate to form an active ternary complex. I am studying the HGF/SF – 

GAG interaction. 

HGF/SF is a multidomain protein which consists of four kringle 

domains (K1-K4) and a N-terminal PAN domain (the N domain). The 

NMR structure of the N domain was solved in 1999 by Zhou et al. and 

the first two domains of HGF/SF, NK1, has been crystallised with and 

without heparin derived oligosaccharides (Lietha et al. 2001). We are 

investigating this interaction further using NMR and other biophysical 

techniques. 

We have expressed the N-terminal domain, the first kringle domain K1 

and the two domains together NK1. We have also prepared a library 

of chondroitin, dermatan and heparin derived oligosaccharides which 

were used in binding studies to the N-domain. Titrations of these GAG 

oligosaccharides were followed by recording a series of 1H-15N HSQC 

spectra. The data shows that the N domain binds with a KD in the low 

uM range to all three GAGs, although heparin is the strongest binder. 

Interestingly, heparin binds to the protein in a two-phase mode. This is 

evidenced by HSQC cross peaks changing the direction of movement 

after reaching 1 : 0.5 protein : ligand ratio. This is likely to be due to the 

formation and breaking of the GAG : protein complex. The NMR data is 

supplemented by ITC, mass spectrometry, affinity chromatography and 

gel filtration studies. 

References 

Lietha, D. et al., 2001. Crystal structures of NK1–heparin complexes 

reveal the basis for NK1 activity and enable engineering of potent 

agonists of the MET receptor. The EMBO Journal, 20(20). 

Zhou, H. et al., 1999. Identification and Dynamics of a Heparin-Binding 

Site in Hepatocyte Growth Factor. Biochemistry, 38(45), 14793-14802. 

Mi26 

NMR characterization of Aβ peptides - tetracycline interaction 

Airoldi, Cristina 1 ; Sironi, Erika 1 ; Colombo, Laura 2 ; Manzoni, Claudia 2 ; 

Natalello, Antonino 1 ; Del Favero, Elena 3 ; Cantù, Laura 3 ; Doglia, Silvia 

Maria 1 ; Nicotra, Francesco 1 ; Salmona, Mario 2 

1 Università degli Studi di Milano-Bicocca, Department of Biotechnology 

and Biosciences, Milan, Italy; 2 Istituto di Ricerche Farmacologiche 

“Mario Negri”, Department of Molecular Biochemistry and Pharmach, 

Milan, Italy; 3 Università degli Studi di Milano LITA, Department of 

Chemistry, Biochemistry and Biotech, Milan, Italy 

Cerebral deposition of Aβ-amyloid is a major neuropathological feature 

in Alzheimer’s disease. It was demonstrated that tetracyclines, classical 

antibiotics, exhibit anti-amyloidogenic activity. This capacity was 

determined by the exposure of Aβ1-40 and Aβ1-42 amyloid peptides 

to the drugs followed by the electron microscopy examination of the 

amyloid fibrils spontaneously formed and quantified with thioflavine 

T binding assay. The drugs reduced also the resistance of Aβ1-40 

and Aβ1-42 amyloid fibrils to trypsin digestion. Tetracyclines not only 

inhibited the Aβ-amyloid aggregates formation but also disassembled 

the pre-formed fibrils. 

Nevertheless, structural information about Aβ peptides and tetracyclines 


interaction is not yet available. In order to elucidate the main features 

of this interaction, we employed NMR spectroscopy, supported by other 

bio-physical techniques, such as atomic force microscopy, electron 

microscopy, FT-IR spectroscopy and laser light-scattering. 

Data collected will be described in this communication. 

Mi27 

A new expression system enabling plastocyanin and 

photosystem I subunit PsaF interaction studies 

Farkas, Daniel; Hansson, Örjan 

University of Gothenburg, Chemistry, Gothenburg, Sweden 

In the photosynthetic electron-transfer chain, plastocyanin is involved 

in shuttling electrons from cytochrome b6f to the photooxidized special 

pair in photosystem I. Structural and mutagenesis studies have revealed 

two negatively charged patches important for the docking and the 

subsequent electron transfer from plastocyanin to photosystem I. The 

high specificity of plastocyanin towards photosystem I found in higher 

plants is believed to be linked to the evolutionary progress of a partially 

lumen-exposed photosystem I subunit called PsaF. This project aims 

to reveal the molecular mechanism behind the interaction between 

plastocyanin and PsaF. 

Regions of special significance for the plastocyanin–PsaF interaction 

can be explored by monitoring the diamagnetic chemical shifts in the 

15N-HSQC spectrum. By utilizing the paramagnetic properties of the 

plastocyanin-bound copper (II), it is believed that enough pseudocontact 

shifts can be gained to model the plastocyanin-PsaF complex. In 

combination with EPR spin-labelling experiments we believe these 

techniques can aid each other to give a more complete description of 

the initial plastocyanin-photosytem I docking event and its dynamics. 

Here we will describe the expression system for PsaF and plastocyanin 

developed by us, as well as preliminary NMR- and EPR-studies. 

Mi28 

TINS: New opportunities for fragment based drug discovery 

Heetebrij, Rob 1 ; AB, Eiso 1 ; Kobayashi, Masakazu 1 ; Loch, Caroline 1 ; Fruh, 

Virginie 2 ; Figaroa, Francis 1 ; Hollander, Johan 1 ; Siegal, Gregg 1 

1 ZoBio, Leiden, Netherlands; 2 Leiden university, Chemistry, Leiden, 

Netherlands 

Fragment based drug discovery (FBDD), the screening of low molecular 

weight compounds as starting points for drug discovery, is gaining 

increased attention because it generates “lead like” compounds and 

has been successfully applied to challenging targets such as proteinprotein 

interactions. We have developed a method we call TINS, for 

target immobilized NMR screening, that is applicable to targets that 

are not possible to obtain in large quantities and/or are insoluble such 

as integral membrane proteins. In TINS, the target is immobilized on a 

solid support. The mixture of compounds to be tested for binding (up 

to 7 at a time) is pumped over the support and binding is detected 

by 1D 1H NMR spectroscopy of the ligands. We have developed 

hardware that allows simultaneous fl owinjection screening of a 

pharmaceutical target and a reference sample. Reference proteins that 

can be used for screening soluble or membrane bound proteins have 

been characterized. The advantage of using a reference is that the 

weak non-specific interactions between small molecules and proteins 

that give rise to false positive signals are removed. The instrument 

can screen a 1,500 compound collection in less than e week in an 

automated manner using as little as 5 nmol of the target. To date we 

have screened a variety of targets, including both soluble and integral 

membrane proteins. In all cases we have been able to immobilize 

proteins with no observable effect on function as measured by binding 

of known ligands or by enzymatic assay, including GPCRs. The hit rate 

thusfar varied between 3-9%. 

Keywords: Ligand screening, NMR, fragment based drug discovery 


Mi29 

In-vivo 2D-TR-NOE interrogates and ranks ligand-receptor 

interactions in living cells 

Mari, Silvia 1 ; Traversari, Catia 2 ; Invernizzi, Chiara 2 ; Bordignon, 

Bordignon 2 ; Renato, Longhi 3 ; Spitaleri, Andrea 1 ; Rizzardi, Paolo 2 ; Musco, 

Giovanna 1 

1 Dulbecco Telethon Institute c/o S. Raffaele Scientific Institute, 

Biomolecular NMR Laboratory, Milan, Italy; 2 MolMed, Milan, Italy; 3 CNR, 

Milan, Italy 

Integrin aVb3 and the membrane-spanning surface protein 

aminopeptidase N (CD13) play a pivotal role in tumour growth and 

metastatic spread, as they are two of the major membrane bound 

receptors highly expressed on the surface of tumour cells during 

angiogenesis. RGD- and NGR-containing peptides, peptidomimetics, 

and conjugated compounds specifically target the tumor vasculature via 

αVβ3 and CD13 recognition, respectively, thus originating novel direct 

acting vascular targeting agents. A crucial contribution to the efficacy 

of these approaches relies on the characterization of receptor-ligand 

molecular interactions in their natural membrane environment. However, 

this is an inherently difficult goal to achieve. 

Here we show that it is possible to apply 2D-TR-NOE techniques directly 

on human cancer cells to prove selective binding of anti-angiogenic 

ligands to structurally characterized and uncharacterized receptors, 

such as αVβ3 and CD13, respectively. 

Using 2D-TR-NOE experiments we investigated the binding of a 

small library of cyclopeptides (CRGDC, CisoDGRC, AcCisoDGRC, 

CDGRC, CNGRC, cyclo(-RGDfV-))onto 2 human cancer cell lines 

differently expressing αVβ3 and CD13, including a melanoma 

(MSR3:αVβ3+CD13- cells) and a non-small lung carcinoma 

(MR300:αVβ3+CD13+ ) cell line, which display different phenotypes 

for CD13 and integrins. Only very small amount of receptors are 

needed to prove binding (in the picomolar range), as it is sufficient that 

the receptor is detectable by FACS analysis. The method allows using 

different cell lines, with different receptors, which can be also silenced 

with siRNA techniques to prove recognition specificity. Most importantly, 

non specific binding can be straightforwardly established by competitive 

binding with stronger ligands. Finally, we show that 2D-TR-NOESY can 

be applied in living tumor cells to screen varying ligands to the same 

receptor, performing competition experiments thus defining an affinity 

ranking of different ligands in a physiological context. 

Mi30 

Structural basis of membrane insertion by the golgi-specific PH 

domain of FAPP1 

Lenoir, Marc 1 ; Coskun, Ünal 2 ; Buschhorn, Sabine 2 ; Simons, Kai 2 ; 

Overduin, Michael 1 

1 University of Birmingham, Institute for Cancer Studies, Birmingham, 

United Kingdom; 2 Max Planck Institute for Molecular Cell Biology and 

Genetics, Dresden, Germany 

The structural mechanisms underlying Golgi membrane recognition 

are still unknown, nevertheless the targeting domains and 

phosphatidylinositol 4-phosphate (PtdIns(4)P) ligand have been 

identified. We have determined the solution structure of the pleckstrin 

homology (PH) domain of the four-phosphate-adaptor protein 1 (FAPP1) 

by NMR spectroscopy, revealing an exposed hydrophobic protrusion that 

penetrates into mixed micelles designed to mimic the Golgi membrane. 

Recognition of PtdIns(4)P and neighboring phospholipid molecules is 

mediated by an extensive network of interactions revealed by NMR. The 

ABSTRACTS POSTER


relative orientation of the micelle and the ligand compared to the FAPP1 

was search by docking methods relying on our NMR data. Hydrophobic 

insertion occurs even in the absence of ligand, with a perpendicular 

orientation of the protein on the micelle based on reduced solvation and 

paramagnetic relaxation enhancement (PRE) of embedded backbone 

and side chain atoms. Our observations also indicated that these 

interactions occur without significant conformational changes. 

Our The NMR results show that the individual phospholipid and PtdIns(4) 

P binding modes are preserved in these micelle complexes. The two 

interactions clearly reinforce each other, involving an extensive binding 

surface which is largely responsible for Golgi membrane targeting by 

the FAPP1 PH domain. 

The conservation of key binding features in related PH domains 

indicates that hydrophobic insertion is the dominant mode of Golgi 

recognition, and may also contribute to the deformation and vesiculation 

of the bilayer as its lipid components are trafficked to the cell surface. 

Mi31 

NMR interaction study between E. coli ribosomal protein S1 and 

the 30S subunit of the ribosome 

Giraud, Pierre; Sizun, Christina; Bontems, François 

CNRS-ICSN-RMN, Gif sur Yvette, France 

The level of gene expression and hence of proteins, is largely dependent 

on messenger RNA concentration. This is ruled by its translation rate 

on the one hand and its degradation rate on the other hand. We are 

interested in the ribosomal protein S1 which is involved in both a priori 

opposite functions: translation initiation and inactivation of mRNAs. 

In prokaryotic cells, like in eukaryotic cells, translation starts with the 

recognition of the initiation codon. However in prokaryotes additional 

signals, like the Shine-Dalgarno (SD) sequence, are needed by the 

ribosome to recognize the start codon among synonymous triplets. 

When the SD sequence is not present or degenerated, ribosomal 

protein S1 is essential for the ribosome to recruit mRNA. The functions 

of S1 are well characterized, but its mode of action has not been 

fully elucidated yet. The overall purpose of the project is to establish 

molecular bases for the activity of S1. 

S1 is a modular protein, composed of a repetition of six homologous 

domains: OB-folds termed “S1 domains”. The two N-terminal domains 

(F1 & F2) form the functional ribosome binding domain F12. The three 

central domains (F3, F4 & F5) form the mRNA-binding core F35. Our 

group has characterized the mRNA-binding site of F35 by chemical 

shift mapping. We have also analyzed interactions between different 

fragments of S1 with each another. 

In parallel we wanted to probe the fixation of S1 to the ribosome. The 

titration of S1 or its F12 domain with the 30S ribosomal subunit showed 

that the two N-terminal domains behave differently. The peak intensities 

corresponding to residues of the second domain F2 decreased more 

rapidly than those of the first domain F1, indicating a fast exchange 

regime for F2. When the F36 fragment was titrated against the 30S 

ribosomal subunit, we observed a line broadening similar to that of F2, 

except for the unstructured C-terminal end. We interpret this in terms of 

two binding modes for S1 to the 30S subunit. A first mode of low affinity 

implies domains F2 to F6. The first domain F1 displays a second fixation 

mode, of higher affinity, which results in intermediate to slow exchange. 

84 

Mi32 

Interaction of the human membrane protein VDAC1 with various 

effectors 

Villinger, Saskia 1 ; Bayrhuber, Monika 1 ; Meins, Thomas 2 ; Habeck, 

Michael 3 ; Becker, Stefan 1 ; Giller, Karin 1 ; Vonrhein, Clemens 4 ; Zeth, 

Kornelius 3 ; Griesinger, Christian 1 ; Zweckstetter, Markus 1 

1 Max Planck Institute for Biophysical Chemistry, Department of NMRbased 

Structural Biology, Göttingen, Germany; 2 Max Planck Institute 

for Biochemistry, Department of Membrane Biochemistry, Martinsried, 

Germany; 3 Max Planck Institute for Developmental Biology, Department 

of Protein Evolution, Tübingen, Germany; 4 Global Phasing Ltd., Sheraton 

House, Castle Park, Cambridge, United Kingdom 

The voltage-dependent anion channel (VDAC) is the most abundant 

protein of the outer mitochondrial outer membrane. VDAC is 

homologous to bacterial porins and forms an aqueous pore with a 

diameter of 2.5-3 nm in planar lipid membranes. The channel is 

responsible for most of the metabolite flux across the membrane, 

therefore regulating the energy metabolism of the cell, and is involved 

in Ca 2+ -signalling. Furthermore, there is substantial evidence that VDAC 

plays an essential role in apoptosis by direct interaction with various 

pro- and anti-apoptotic proteins. The functional mechanism, however, is 

still a matter of debate. There is considerable evidence for the existence 

of several conformations of VDAC. In the open conformation (at low 

potentials) the channel is highly permeable for ATP, ADP and other 

large anions. In contrast, at high positive or negative potentials VDAC 

closes, inverting its selectivity and preferring small cations like Ca 2+ . 

Additionally, VDAC is capable of oligomerisation and is proposed to be 

a major component of the “permeability transition pore” that in its open 

state promotes apoptotic cell death by the release of cytochrome c to 

the cytosol. 

By the combination of solution NMR spectroscopy and X-Ray 

crystallography (accomplished by collaboration with the group of 

Dr. Kornelius Zeth, MPI for Evolutionary Biology, Tübingen) we have 

solved the structure of the most prominent human isoform VDAC1 

in detergent micelles. NMR spectra showed a quality sufficient for 

backbone resonance assignments of up to approximately 70% (~90% 

of β-barrel residues). We have performed interaction studies of VDAC1 

with several effectors, including the natural transport substrate ATP, as 

well as Gd 3+ and ruthenium red, which are both supposed to bind to 

Ca 2+ -binding sites. Structural and functional implications can be inferred 

from the observed chemical shift changes and paramagnetic relaxation 

enhancement. 

Mi33 

Real-time monitoring of transaminase reactions using DNP- 

NMR 

Karlsson, Magnus 1 ; Jensen, Pernille R. 1 ; Meier, Sebastian 2 ; Duus, Jens 

Ø. 2 ; Lerche, Mathilde H 1 

1 Imagnia AB, Malmö, Sweden; 2 Carlsberg Laboratory, Valby, Denmark 

The DNP-NMR technique combined with 13 C labeled metabolic 

substrates provides signal enhancement sufficient for real time 

monitoring of several types of enzymatic reactions. One important class 

of metabolic reactions possible to monitor by DNP-NMR is catalyzed by 

transaminases. These enzymes catalyze conversion of an amino acid 

to its keto analogue or vice versa, linking metabolic substrates such 

as pyruvate to alanine, oxaloacetate to aspartate and ketoglutarate to 

glutamate. Since many of the transaminase reactions have a K eq close 

to 1, either the amino acid or the keto-acid can be used as a DNPsubstrate. 

While it is fairly straightforward to prepare DNP samples of 

amino acids in low concentrations, certain applications (e.g. in vivo 

experiments) calls for highly concentrated substances which demands 

special preparation procedures. Here we describe two preparation 


methods which make it possible to achieve highly concentrated 

solutions of hyperpolarized 13 C labeled amino acids. We also describe 

a relaxation phenomenon which affects the amino acids in solutions 

above neutral pH. 

Paramagnetic 


Pa10 

An improved paramagnetic tag for proteins 

Peters, Fabian; Leonov, Andrei; Schmidt, Holger; Becker, Stefan; 

Griesinger, Christian 

Max Planck Institute for Biophysical Chemistry, NMR based Structural 

Biology, Göttingen, Germany 

Residual dipolar couplings (RDCs) are useful NMR parameters for 

structure determination of proteins since they contain information about 

relative orientations of internuclear vectors. Paramagnetic tagging is 

of special importance when it comes to the determination of motions 

of domains with respect to one another since conventional external 

alignment media fail to report on these domain motions [1,2]. 

Several paramagnetic tags have been developed in the past; however, 

they have failed to yield reliable dipolar couplings in many instances. 

Here we introduce a new nonadentate paramagnetic tag that was 

designed and successfully synthesized. It is attached via a single 

cysteine side chain to the protein. It fulfills 8−9 coordination sites of 

the lanthanide ion thus reducing spurious interactions of the tag with 

charged side chains of the protein. The lanthanide complex forms a 

single stereoisomer, which is a requirement for working with larger 

proteins. We obtained RDCs up to 15 Hz for the backbone amides at 

900 MHz with Tb 3+ . 

Results on the proteins to which the tag was attached will be reported. 

[1] F. Rodriguez-Castaneda, P. Haberz, A. Leonov, C. Griesinger, Magn. 

Res. Chem. 2006, 44, 10−16. 

[2] P. Haberz, F. Rodriguez-Castaneda, J. Junker, S. Becker, A. Leonov, 

C. Griesinger, Org. Lett. 2006, 8, 1275−1278. 

Pa11 

Location of transition-metal ions within inorganic frameworks: 

investigation by solid-state NMR and first principles 

calculations 

Mali, Gregor; Kauèiè, Venèeslav 

National Institute of Chemistry, Ljubljana, Slovenia 

Many new inorganic materials contain transition-metal ions. In our 

laboratory we are investigating transition-metal modified micro- and 

mesoporous silicates and aluminophosphates, and transition-metal 

lithium silicates (Li 2 MnSiO 4 , Li 2 FeSiO 4 , Li 2 CoSiO 4 , ...) The former group 

of materials represents potential catalysts for cleaner oxidations, and 

the latter series of materials could find their use in the field of lithium 

batteries. 

In this contribution we investigate, what combined use of solid-state 

NMR spectroscopy and first-principles DFT calculations can tell about 

the location of transition-metal ions within inorganic frameworks. 

Because transition-metal ions are predominantly paramagnetic, they 

substantially influence NMR spectra of nearby nuclei ( 31 P, 29 Si, 6 Li, 7 Li 

etc.). Their influence is reflected in spectra in two ways, (i) direct dipolar 

interaction of unpaired electronic spins with nuclear spins induces 

substantial broadening of NMR signals, (ii) delocalization of unpaired 

spins over covalent bonds results in strong hyperfine couplings of these 

spins with neighbouring atomic nuclei, which is further manifested 

through large shifts of NMR signals. 

In the course of our study we recorded phosphorus NMR spectra of 

Mn-, Fe-, Co- and Ni-modified aluminophosphate AlPO 4 -34 and silicon 

spectra of several Mn-modified micro- and mesoporous silicates. We 

also inspected lithium spectra of several polymorphs of Li 2 MnSiO 4 . Then 

we carried out first-principles calculations for corresponding 3D periodic 

ABSTRACTS POSTER


systems and tried to relate the calculated transferred spin-density to 

measured hyperfine-coupling induced shifts of NMR signals. In this way 

we were able to obtain an insight into location of transition-metal ions 

within the studied materials. 

Pa12 

Lanthanide paramagnetic probes for NMR studies of molecular 

conformational dynamics and MRI temperature monitoring 

Babailov, Sergey 

Nikolaev’s Institute of Inorganic Chemistry, Structural Chemistry, 

Novosibirsk, Russian Federation 

Information on the molecular structure of coordination compounds of 

Ln in solutions underlies the current technologies for pharmaceutical 

preparations for photodynamic therapy, contrast reagents for magnetic 

resonance tomography and sensors for biology and medicine [1]. 

The following peculiarity of using DNMR for studies of paramagnetic 

Ln compounds deserves special attention. As a result of lanthanideinduced 

shifts in paramagnetic Ln complexes, the range of measurable 

rate constants expands considerably compared to the analogous range 

in diamagnetic compounds. This may be illustrated by reference to a 

degenerate two-site exchange (1). For an instrument with an operating 

frequency for protons of 500 MHz and assuming a value of Δν=50 

KHz (range of shifts is 100 ppm) in a paramagnetic Ln compounds and 

having an error of half-width (W*) determination of ~ 1 Hz, the largest 

value of a rate constant that can be measured is k(max) ~ 10 GHz. 

This is much large than the value ~ 1 MHz measurable in diamagnetic 

compounds (Δν~ 500 Hz). 

k(max) ~ 0.5(pi)( Δν )(Δν )/(W* - 1/((pi)T)) (1). 

T is spin-spin relaxation time. Thus, using the paramagnetic lanthanide 

probe method for investigation of intramolecular dynamics of ligands 

coordinated to this lanthanide cation is equivalent to the use of a 

NMR spectrometer with an operating frequency of 10 GHz, which is 

unavailable for present instruments. Therefore, the potential of this 

method for studying the structures and intramolecular dynamics of 

various ligands complexes with paramagnetic lanthanide cations is 

higher than in the case of complexes with diamagnetic cations. It is 

illustrated by several systems. 

The high stability of the [Ln(EDTA)] anion in aqueous solvents makes it 

possible to use these compounds as thermometric NMR sensors directly 

in reaction media (for in situ monitoring over temperature) and in MRI 

for 3D mapping of the temperature distribution of an animal body. 

The study was carried out under financial support of the Presidium of 

the Siberian Division of the Russian Academy of Sciences (grant N 67). 

1. Babailov S.P., Progr. Nucl. Magn. Res. Spectrosc., 2008, 52, N 1, P. 1 

Pa13 

Hyperfine shifts in NMR spectra of macrobicyclic tris-dioximate 

complexes with encapsulated cobalt(II) ion: a new class of 

paramagnetic tags 

Novikov, Valentin; Lebedev, Andrey; Belov, Alexander; Voloshin, Yan 

Nesmeyanov Institute of Organoelement Compounds RAS, Moscow, 


Hyperfine shifts in NMR spectra of paramagnetic ion’s complexes 

provide a wealth of information about structure and dynamics of 

investigated molecules [1]. The paramagnetic tags, containing 

lanthanide of d-group metal ions, are nowadays broadly used for 

structural characterization of complex biological system [2]. The 

cage complexes with encapsulated cobalt(II) ion [3] are perspective 

paramagnetic labels due to complete isolation of encapsulated 

paramagnetic ion from the media and, hence, stability of the complex 

86 

and independence of its magnetic properties on the media properties. 

The 1 H NMR spectra of C 16 H 33 -substituted cobalt(II) clatrochelate provide 

an example of significant pseudo-contact shifts, leading to a complete 

resolution of the fifteen signals of methylene group protons, otherwise 

heavily overlapped. 

The possible functionalization by six ribbed and two apical substituents 

provide the rich capabilities to fine-tune the properties of encapsulated 

ion to achieve the properties desired. The spin-transition behavior, 

observed for some complexes, is also of a great interest. 

Acknowledgements. 

This work was supported by the Zamaraev International Charitable 

Scientific Foundation. 

References. 

1. I. Bertini, C. Luchinat, G. Parigi, Solution NMR of paramagnetic 

molecules - applications to metallobiomolecules and models, Current 

Methods in Inorganic Chemistry, vol.2, Elsevier, Amsterdam, 2001. 

2. R.R. Sharp, Nuclear Magnetic Resonance, Volume 34, 553-596. 

3. Ya.Z. Voloshin, O.A. Varzatsky, Yu.N. Bubnov, Russian Chemical 

Bulletin, 2007, 56, 577-605. 

Pa14 

Conformational distribution in paramagnetic labeling of proteins 

Polyhach, Yevhen; Bordignon, Enrica; Jeschke, Gunnar 

ETH Zürich, Lab. Phys. Chem., Zürich, Switzerland 

Spin labels can be considered as paramagnetic markers attached to 

a biomacromolecule via a linker. The linker must strike a particular 

compromise between rigidity that ensures a precise positioning of the 

paramagnetic marker and flexibility that allows for labeling without 

distortion of the native structure of the biomacromolecule. While quite 

rigid labels can be accommodated by DNA and RNA, this is usually not 

the case for proteins. The most frequently used methanethiosulfonate 

spin label (MTSSL) has as many as five side chain dihedral angles 

that can adopt two or three values. If this conformational space is not 

constrained by interactions with neighboring side groups, the label 

position has a mean square deviation of 6.3 Å, corresponding to a 

B-factor of 500 Å 2 . Usually the conformational space is considerably 

constrained by interaction of the label with the protein. Fast and 

complete sampling of this space can be achieved by a rotamer 

library approach.[1] This contribution describes such an approach, 

including derivation of the rotamer library, modeling of the label-protein 

interaction, tests on known structures, and applications in structure 

determination by EPR distance measurements. Examples are provided 

for MTSSL in sodium/proton antiporter NhaA of Escherichia coli and 

for the iodacetamido-PROXYL spin label in major plant light harvesting 

complex IIb of garden pea (Pisum sativum). 

[1]G. Jeschke, Ye. Polyhach, Phys. Chem. Chem. Phys., 9, 1895-1910 

(2007), Distance measurements on spin-labelled biomacromolecules by 

pulsed electron paramagnetic resonance 

Pa15 

Paramagnetic relaxation in unfolded proteins: theory and 

application to drkN SH3 and ubiquitin 

Xue, Yi; Podkorytov, Ivan S; Rao, D Krishna; Benjamin, Nathan; Sun, 

Honglei; Skrynnikov, Nikolai R 

Purdue University, West Lafayette, United States 

Site-directed spin labeling in combination with paramagnetic relaxation 

enhancement (PRE) measurements is one of the most promising 

techniques for studying unfolded proteins. Since the pioneering work 

of Gillespie and Shortle (J Mol Biol 1997;268:158), PRE data from 

unfolded proteins have been interpreted using the theory that was 


originally developed for rotational spin relaxation. At the same time, it 

can be readily recognized that the relative motion of the paramagnetic 

tag and the spin probe (1HN) is best described as a translation. With 

this notion in mind, we developed a number of models for the PRE 

effect in unfolded proteins: (i) mutual diffusion of the two tethered 

spheres, (ii) mutual diffusion of the two tethered spheres subject to a 

harmonic potential, (iii) mutual diffusion of the two tethered spheres 

subject to a simulated mean-force potential (Smoluchowski equation); 

(iv) explicit-atom molecular dynamics simulation. The theory has been 

tested by measuring the PRE rates in three MTSL-tagged mutants 

of drkN SH3 domain (2M guanidinium chloride, pH 6) and three 

MTSL-tagged mutants of ubiquitin (8M urea, pH 2). Two modifications 

introduced into the measurement scheme—using a reference 

compound to calibrate the signals from the two samples (oxidized and 

reduced) and using peak volumes instead of intensities to determine 

the PRE rates—lead to a substantial improvement in the quality of 

data. The data from the denatured drkN SH3 point toward a presence 

of compact (collapsed) species. At the same time, the data from the 

denatured ubiquitin are consistent with the random coil model, in 

agreement with the previous observations by Wirmer, Peti, and Scwalbe 

(JBNMR 2006;35:175). Currently, the analysis is extended using MD 

simulations in implicit solvent and backbone 15N relaxation data with 

the goal to obtain a comprehensive picture of internal dynamics in 

unfolded proteins. 

Pa16 

NMR relaxation induced by paramagnetic particles 

Gossuin, Yves; Gillis, Pierre; Vuong, Quoc Lam 

University of Mons, Biological Physics Department, Mons, Belgium 

Rare-earth oxide and hydroxide nanoparticles have been developed 

by several groups, in order to produce a new generation of Magnetic 

Resonance Imaging (MRI) contrast agents 1-4 . However, the proton 

relaxation induced by these compounds is not well described in the 

literature. To evaluate their potential as MRI contrast media, it is necessary 

to understand the relaxation of water protons in the presence of these 

paramagnetic particles. 

The transverse relaxation rates of dysprosium oxide, gadolinium hydroxide, 

holmium oxide and erbium oxide solutions have been measured at 

different magnetic fields, from 0.23 T to 11.7 T. The influence of the 

echo-time on the transverse relaxation was also evaluated for the different 

compounds. The observed dependence of 1/T 2 with the magnetic field 

and the influence of echo-time were then confronted with the predictions 

of different relaxation models, including the well-know outer sphere theory 

(OS) and static dephasing regime (SDR). 

Except for gadolinium hydroxide particles, which exhibit a proton exchange 

relaxation mechanism, the relaxation induced by paramagnetic particles is 

due to the diffusion of protons near the particles. At low field, OS theory is 

still valid, while at high field the SDR must be used. 

Computer simulations of the transverse relaxation induced by the diffusion 

of water protons nearby paramagnetic particles (for different magnetic 

fields and particle sizes) complete this analysis. It also allows predicting 

the relaxivity of paramagnetic particles at any magnetic field which is 

essential for an application in MRI. 

1. Fortin M-A, Petoral RM, Söderlind F, Klasson A, Engström M, Veres T, 

Käll P-O, Uvdal K. Nanotechnology. 2007; 18: 085608. 

2. Engström M, Klasson A, Pedersen H, Vahlberg C, Käll P-O, Uvdal K. 

Magn Reson Mater. 2006; 19: 180-186. 

3. Bridot J-L, Faure A-C, Laurent S, Rivière C, Billotey C, Hiba B, Janier 

M, Josserand V, Coll J-L, Vander Elst L, Muller RN, Roux S, Perriat P, 

Tillement O. J Am Chem Soc. 2007; 129: 5076-5084. 

4. Petoral RM, Söderlind F, Klasson A, Suska A, Fortin MA, Abrikossova 

N, Selegård L, Käll PO, Engström M, Uvdal K. J Phys Chem C. 2009; 

113: 6913–6920. 

Relaxation 


Re10 

Molecular basis of conformational changes observed in the D1 

domain of annexin 1 from NMR relaxation dispersion and pH 

titration experiments 

Gallopin, Matthieu 1 ; Thureau, Aurelien 2 ; Guittet, Eric 2 ; vanHeijenoort, 

Carine 2 

1 CEA, Orsay, France; 2 CNRS-ICSN, Gif sur Yvette, France 

We have employed transverse relaxation dispersion methods, which 

allow a quantitative measurement of microsecond to millisecond 

motions along proteins, coupled to circular dichroïsm and pH titration 

followed by NMR to analyze the diverse exchange processes occurring 

in domain 1 of human annexin I. Reliable thermodynamic parameters 

ΔG, ΔH and ΔCp are obtained from meticulous analysis of CD curves 

showing a high stability over a large pH range (from 4.0 to 9.0). 

At lower pH, the stability drops rapidly. We used CPMG relaxationcompensated 

dispersion experiments at various pH values from 2.2 

to 9.0 to investigate µs-ms timescale dynamics of D1 under nondenaturing 

conditions. The results uncover two distinctive dynamical 

behaviours at pH equal to or higher than 3.5 and lower than 3.5. 

At pH below 3.5, a uniform exchange contribution in the intermediateslow 

regime is evidenced along the whole sequence of D1. The 

minor conformer is shown to be the unfolded state. A pH titration of 

the sidechain carboxyl group of the Asp and Glu residues permits to 

determine the pKa for the folded states. Asp15 exhibits a very low pKa 

and the results of the relaxation dispersion experiments show that the 

acid unfolding of D1 is coupled to the protonation of Asp15 that disrupts 

a salt bridge with Arg40. 

At pH higher than 4.0, D1 exhibits a fast exchange process at a 

timescale around 500µs localised principally in the B and E helices and 

the C-terminal end of the D helix. The results unravel the existence of, 

at least, two exchanging conformers in the folded state of D1, whose 

differences are located at the interface between helices D1 and D4. 

The data presented here along with bioinformatics informations support 

the proposal that annexin domains have evolved to maintain a key 

interaction between conserved residues that drives its fast and highly 

cooperative folding which is essential for the D1 folding as an initial 

step occurring before the global folding of this multi domain protein. 

Re11 

Probing the conformational entropy of binding to Gal3 using 

different synthetic ligands 

Diehl, Carl 1 ; Genheden, Samuel 2 ; Ryde, Ulf 2 ; Nilsson, Ulf 3 ; Leffler, 

Hakon 4 ; Akke, Mikael 1 

1 Lund University, Biophysical Chemistry, Lund, Sweden; 2 Lund University, 

Theoretical Chemistry, Lund, Sweden; 3 Lund University, Organic 

Chemistry, Lund, Sweden; 4 Lund University, MIG, Lund, Sweden 

The binding of a ligand to a protein can be characterized 

thermodynamically by the free energy of binding (ΔG). The free 

energy can be separated into two parts according to the classical 

thermodynamical equation ΔG = ΔH – TΔS. In structure-based drug 

design the interaction energy (enthalpy) is estimated using molecular 

force fields and high-resolution structures, but the entropic part of the 

interaction typically remains unaccounted for. Using NMR relaxation 

experiments it is possible to probe the probability distribution of 

conformational fluctuations, which is directly related to entropy. 

Here we present model-free order parameters for Galectin-3 in the apo 

state as well as in complex with three different but structurally similar 

ligands with varying dissocation constants. Galectin-3 (Gal3) is a member 

ABSTRACTS POSTER


of a protein family defined by its carbohydrate recognition domain and 

affinity for β-galactosides. Gal-3 is both an intra- and extracellular protein, 

which has been shown to play important roles in inflammation and cancer. 

We measured the 15N longitudinal (R 1 ) and transverse (R 2 ) relaxation rate 

constants and 1 H- 15 N cross relaxation rate constants (NOE) at a static 

magnetic field of 14.1 T. Data were acquired for the backbone amides, 

side-chain arginine Nε and side-chain tryptophane Nε. Based on these 

data we calculated model-free order parameters for each of the four 

different states. 

The four different states show highly similar patterns of order parameters. 

However, there are small but significant changes in the order parameters 

between the different states. The apo state presents the highest order 

parameters of all states, indicating that the backbone of Gal3 becomes 

more flexible upon ligand binding. Yet, the order parameters show an 

increasing trend with decreasing dissociation constants, where lactose 

has the lowest order parameters and the tightest binder has the highest 

order parameters. 

Order parameters calculated from molecular dynamic simulations of 

Galectin-3 in the apo-state and the three different complexes agree well 

with fitted order parameters for both backbone and side-chains. 

Re12 

Field dependent relaxation and polarization transfer processes 

in solid state systems: recent theoretical progress 

Kruk, Danuta 1 ; Fujara, Franz 2 ; Privalov, Alexei 2 ; Medycki, Wojciech 3 

1 Jagiellonian University, Institute of Physics, Cracow, Poland; 2 Technical 

University of Darmstadt, Darmstadt, Germany; 3 Institute of Molecular 

Physics, Poznan, Poland 

A theory of polarization transfer and field dependent relaxation in solid 

state systems, containing mutually coupled spins of spin quantum 

numbers I=1/2 (spins 1/2) and S>=1 (possessing quadrupole 

moments) [1-5], is presented. A proper analysis of relaxation and 

polarization transfer experiments for multi-spin systems is very 

demanding from a theoretical point of view. In such cases averaged 

quadrupolar interactions contribute to the energy level structure of the 

system. In addition, local fluctuations of the electric field gradient tensor 

provide a relaxation mechanism for the S>=1 spin nuclei, affecting 

the I spin relaxation via the mutual I-S dipole-dipole coupling. In 

consequence the commonly used relaxation formulas [6,7] break down, 

leading to serious misinterpretations if applied incorrectly. 

An analysis of the polarization transfer pattern (i.e. the I=1/2 spin 

magnetization measured versus magnetic field) gives information on 

the quadrupolar parameters. Thus, polarization transfer and relaxation 

experiments provide complementary and unique information if properly 

evaluated. 

The presented advanced treatment is very useful for numerous field 

cycling investigations of dynamic processes in various molecular 

systems. The goal of this lecture is to popularize this proper theoretical 

approach with the intention to establish it as a standard tool for analysis 

relaxation data for complex molecular systems. 

[1] D. Kruk, Theory of Evolution and Relaxation of Multi-spin Systems, 

Abramis Academic, Arima Publishing UK, 2007. 

[2] D. Kruk, O. Lips, J. Magn. Reson. 179, (2006) 250-262. 

[3] D. Kruk, O. Lips, SSNMR 28 (2-4), (2005) 180-192. 

[4] D. Kruk, J. Altmann , F. Fujara , A. Gadke, M. Nolte, A.F. Privalov, J. 

Phys. Condens. Matter 17 (3), (2005) 519-533. 

[5] D. Kruk, F. Fujara, P. Gumann, W. Medycki, A. Privalov, C. Tacke, 

SSNMR- in press 

[6] N. Bloembergen, E.M. Purcell, R.V. Pound, Phys. Rev. 73, (1948) 

679-712. 

[7] N. Bloembergen, L.O. Morgan, J. Chem. Phys. 34, (1961) 842-850. 

Acknowledgment - This work has been supported by Grant N N202 

172135 of Polish Ministry of Science and High Education. 

88 

Re13 

Combining NMR diffusion and relaxation measurements to gain 

insight into the supramolecular structure of a pore forming 

cyclic lipodepsipeptide 

Sinnaeve, Davy 1 ; Kieffer, Bruno 2 ; Martins, José C. 1 

1 Ghent University, Organische Chemie, Gent, Belgium; 2 Ecole Supérieure 

de Biotechnologie de Strasbourg, Illkirch, France 

The potential of diffusion and heteronuclear relaxation measurements 

to address supramolecular structure formation and shape has been 

investigated using pseudodesmin A[1]. This is a small natural cyclic 

lipodepsipeptide (CLP) that belongs to a class of compounds that is 

known to be able to form ion pores in cellular membranes. We have 

shown through translational diffusion NMR that the compound is able to 

self-associate to very large structures in apolar organic solvents such 

as chloroform. The solution structure derived in non-self associating 

conditions (polar solvent, acetonitrile) is a short amphipatic left-handed 

helix. This structure suggests a model for the self-association where 

the helices stack upon each other, while the hydrophilic side of the 

molecules pack together to minimize the hydrophilic contact surface, 

effectively creating a hydrophilic tunnel that is able to traverse the 

membrane. To validate our proposed model, heteronuclear 13 Cα 

relaxation (T 1 , T 2 , 1 H-{ 13 C}-nOe, relaxation dispersion) measurements 

were performed at both 500 MHz and 700 MHz on both the monomeric 

(acetonitrile) and different concentrations of the oligomeric state 

(chloroform). Some observed deviations from the theoretically expected 

correlation between T 1 , T 2 and nOe values as well as among the two 

magnetic fields can be explained by dissimilar ensemble averaging of 

these different parameters over all oligomer sizes as is shown through 

simulation. Confrontation of the relaxation data with the monomer 

solution structure shows a clear dependence of the CH bond vector 

orientation which increases with higher concentration, providing 

evidence of an increasingly anisotropic supramolecular structure of 

increasing length but constant diameter. In addition, the orientation of 

the experimental rotational diffusion tensor reveals the main direction 

of stacking to be parallel with the helix structure, in agreement with 

our proposed model. The information thus obtained will prove to be 

invaluable to guide the elucidation of the molecular details of the 

supramolecular structure using intermolecular nOe contacts. 

[1] Sinnaeve et al, Tetrahedron, in press, DOI: 10.1016/j. 

tet.2009.03.045 

Re14 

Characterization of crude oil products by low field NMR 

Maddinelli, Giuseppe 1 ; Pavoni, Silvia 2 ; Passerini, Cinzia 2 

1 ENI, Physical Chemistry Department, San Donato, Milan, Italy; 2 ENI, 

Downstream Technology Department, San Donato, Milan, Italy 

Benchtop and mobile NMR Relaxometers are now becoming very 

popular because of their great flexibility in application to materials 

studies. Single-sided NMR devices are especially interesting, because 

of their unique capability to extend analysis to materials which cannot 

fit into the bore of standard NMR magnets. These instruments are now 

comparable to sophisticated spectrometers allowing the application 

of complex sequences even for MRI experiments. In this study we 

have applied a benchtop NMR analyzer (20 MHz) equipped with 

field gradients capabilities inside a permanent magnet and with a 

commercial single-sided NMR device working at 15.8 MHz. NMR pulsed 

field gradient diffusion experiments, relaxation times and hydrogen 

content measurements were applied to collect information on the 

physical and chemical properties of several crude oils becoming from 

very different reservoirs. The results were compared with rheological 

and chemical parameters obtained by standard ASTM methods. An 


appreciable correlation between NMR parameters, such as diffusion 

coefficients and transverse proton relaxation times, and material 

properties (e.g. viscosity and API gravity) was found. The results 

obtained with the different tools (volume and surface measurements) 

are examined and discussed. The relaxation times distribution 

spectra obtained by applying an Inverse Laplace Transformation (ILT) 

algorithm, give a substantial fingerprint of the type of oil useful to 

evaluate the type of chemical components present in such a complex 

hydrocarbon mixture. Moreover, a correlation between the relaxation 

times distribution analyzed by a statistical method and the asphaltene 

content was found. This method could provide a useful determination of 

the heavy chemical component contained in the crude oils. A different 

approach based on the direct analysis of the CPMG decay curves by 

applying a neural network algorithm, confirmed the results and provided 

other correlations. 

The experimental data demonstrate the feasible application of such 

methods on a large range of samples, providing an efficient and fast 

method suitable to be applied directly on the industrial site. 

Re15 

Proton tunnelling in the hydrogen bond: 18 O isotope effects of 

the heavy atom framework 

Frantsuzov, Ilya; Horsewill, A. J. 

University of Nottingham, School of Physics and Astronomy, Nottingham, 


Proton transfer is one of the simplest chemical reactions and is 

therefore of fundamental interest to all processes which involve the 

motion of atoms over a potential energy surface (PES). There has been 

much interest in concerted double proton transfer in the carboxylic 

acid dimers where the barrier-crossing process at low temperature 

is dominated by incoherent quantum tunnelling. One of the simplest 

members of this family, benzoic acid, is established as the model 

system for translational atomic tunnelling. [1] 

We have shown over a number of years how field-cycling NMR 

relaxometry can be employed to map out the spectral density 

associated with the proton transfer motion and provide an accurate 

measure of the proton tunnelling rate and its temperature dependence. 

It is well known in quantum mechanics that the tunnelling phenomenon 

is exponentially sensitive to the square root of the particle mass; 

isotopic substitution of 1 H with 2 H gives rise to a reduction in the 

tunnelling rate by a factor of approximately 300. 

The proton transfer coordinate involves displacements of the carbon 

and oxygen atoms as well as hydrogen. The motion is therefore properly 

described as motion on a multi-dimensional PES and is that of a quasiparticle 

with mass dressed by small admixtures of the heavy atoms. We 

report on a systematic investigation to explore the effects of isotopic 

substitution of oxygen. The spectral density in 16 O and 18 O samples has 

been compared using field-dependent measurements of the 1 H T 1 by 

field-cycling NMR. An isotope effect of order 15% is observed. 

References 

[1] A.J. Horsewill, Progr. Nucl. Magn. Reson. Spectrosc. 52 (2008) 170- 

196 


Re16 

Unite experiment and simulation: structure and dynamics of a 

regulatory RNA from Hepatitis B virus 

Petzold, Katja 1 ; Lindorff-Larsen, Kresten 2 ; Zdunek, Janusz 3 ; Varnai, 

Peter 4 ; Vendruscolo, Michele 5 ; Schleucher, Jürgen 1 

1 Umeå University, Medical Biochemistry and Biophysics, Umeå, Sweden; 

2 University of Copenhagen, Structural Biology and NMR Laboratory, 

Department of Molecular Biology, Copenhagen, Denmark; 3 Protein 

Constructor Developers company, Umeå, Sweden; 4 University of Sussex, 

Department of Chemistry, Sussex, United Kingdom; 5 University of 

Cambridge, Department of Chemistry, Cambridge, United Kingdom 

The „epsilon“ RNA of the Hepatitis B Virus (HBV) is a central player 

in the viral life cycle, because it is recognized by the viral reverse 

transcriptase and triggers so encapsidation and production of new viral 

DNA. We have previously determined the structure and analyzed the 

dynamics of the apical loop of epsilon by NMR (Flodell et al NAR 2006 

& Petzold et al., NAR 2007). Dynamics was studied using both C-H and 

H-H vectors as probes, yielding a detailed picture of the RNA’s motions. 

Distinct patterns of motion were observed for several fully conserved 

nucleotides, which strongly suggests that motion of these nucleotides 

plays a role for the RNA’s biological function. Because motion is not 

captured in static structures, a static structure represents at best a 

snapshot of the biologically relevant structural ensemble of the RNA. 

Therefore we are calculating a dynamic structural ensemble of the 

RNA by restrained Molecular Dynamics (MD) (Lindorff-Larsen et al. 

Nature 2005). These MD simulations use structural and motional data 

as restraints (order parameter and NOE), yielding a structural ensemble 

which represents structural AND dynamical properties combined. 

The so created RNA dynamics structural ensemble better represents 

mobile nucleotides, and improves the agreement between experimental 

and simulated structural data. Using this combined experimental and 

simulational approach, we derive high-resolution motional models. 

These models describe the conformational space sampled by the RNA, 

and recognized via conformational capture by the reverse transcriptase. 

The conformational space also defines the conformations to be 

considered in the development of novel drug classes which target RNA 

structures. 

Re17 

Non-linearity, frequency shifts and other surprises with nuclear 

spin noise 

Mueller, Norbert 1 ; Nausner, Martin 1 ; Schlagnitweit, Judith 1 ; Jerschow, 

Alexej 2 ; Smrecki, Vilko 3 

1 Johannes Kepler University, Linz, Austria; 2 New York University, New 

York, United States; 3 Rudjer Boskovic Institute, NMR-center, Zagreb, 

Croatia 

We have investigated spin noise spectra of liquids and solids under a 

variety of conditions to put some theoretical predictions to experimental 

test. It was found that the observed line shapes of the proton nuclear 

spin noise spectra depend in complex ways on the properties 

of the resonance circuit, the spin density, transverse relaxation, 

inhomogeneous broadening, radiation damping and temperature. 

Although spin noise spectra by their nature are power or magnitude 

spectra, they generally exhibit dispersion-like line shapes. According 

to theory [1] spin noise signals should appear as negative deviations 

from the otherwise flat positive (thermal) noise power baseline, if 

the tuning frequency of the circuit is equal or close to the Larmor 

frequency. However, we find this predicted symmetrical “dip” line 

shape at a considerable tuning offset, the “spin noise tuning optimum” 

(SNTO). At the conventional tuning optimum, which can be several 

kilohertz away from the SNTO, a dispersion like line shape is usually 

observed. Systematic investigations revealed that the peak areas are 

ABSTRACTS POSTER


not proportional to the number of nuclei. For instance broad OH proton 

signals appear much larger than narrow signals of methyl groups. 

We have found semi-quantitative explanations of these experimental 

observations, which also include frequency shifts at high spin densities 

and dependence of line shapes on sample and coil temperatures. There 

is a complex interplay of transverse relaxation, radiation damping, 

and inhomogeneous broadening which can be tracked by numerical 

simulations. [2] 

Acknowledgements of financial support by: Austrian Marshall Plan 

Foundation (Scholarship to M.N.), Croatian Ministry of Science, 

Education and Sports (project 098-0982929-2917, to V.S.), US National 

Science Foundation (CHE-0550054, to A.J.), Ã–AD (WTZ AT-HR 

19/2008, to N.M.), Austrian Science Funds FWF (P19635-N17 to N.M.) 

References: 

[1] M.A. McCoy, R.R. Ernst (1989) Chem. Phys. Lett. 139, 587 [2] 

M. Nausner, J. Schlagnitweit, V. Smrecki, X. Yang, A. Jerschow and 

N. Mueller (2009) J. Magn. Reson., Article in Press doi:10.1016/j. 

jmr.2009.01.019 

Re18 

Low-field NMR of heterogeneous mixtures and polymers under 

stress 

Böhme, Ute; Scheler, Ulrich 

Leibniz Institute of Polymer Research Dresden, Dresden, Germany 

Small and low-cost NMR systems based on permanent magnets provide 

an interesting alternative for NMR in materials characterization. They 

are used as tools for the routine analysis of foodstuffs, cosmetics 

and chemicals. They are of particular interest, when chemical-shift 

information is not required, i.e., when there is apriory knowledge about 

the system. NMR relaxation times provide a wealth of information 

on molecular mobility, they are influenced by the composition and 

treatment of the sample. Impacts like stress, heating or cooling change 

the molecular mobility. Extending the relaxation time measurements 

to two-dimensions significantly enhances the resolution of the 

components of the sample despite lack of chemical information. For our 

investigations we use an in-house built portable NMR system based on 

a Halbach arrangement of permanent magnets with a proton Larmor 

frequency of 32 MHz and a probe head with a solenoid coil. 

Examples shown include the determination of the principal components 

of water- in- oil- emulsion and stretched polypropylene using T1-T2 

correlation experiments at low field. 

The extension to two dimensions faciltates the assignments in the 

emulsion, which are confirmed by high-field measurements extending 

to chemical shift as a third dimension. 

The study of polypropylene under stress confirms changes in the 

polymer orientation which relax after the stress is released. 

Re19 

Reorientational and internal dynamics of a disaccharide 

molecule described by a novel model on the basis of NMR 

relaxation data 

Zerbetto, Mirco 1 ; Kotsyubynskyy, Dmytro 2 ; Polimeno, Antonino 1 ; 

Ghalebani, Leila 2 ; Kowalewski, Jozef 2 ; Meirovitch, Eva 3 

1 Università degli Studi di Padova, Dipartimento di Scienze Chimiche, 

Padova, Italy; 2 Stockholm University, D. of Physical, Inorganic and 

Structural Chemistry, Stockholm, Sweden; 3 Bar-Ilan University, The Mina 

& Everard Goodman Fac. of Life Sciences, Ramat-Gan, Israel 

The description of the reorientational dynamics of flexible molecules is 

challenging. Exact numerical treatment of the comprehensive - overall 

and internal - dynamics is essentially impossible, in particular when the 

90 

rates of these motions are comparable. Simple models based on modedecoupling, 

such as model-free and various jump models, are likely not 

to be useful because they will be used outside of their validity range. 

An approach which does account rigorously for mode-coupling, and 

other important aspects omitted in the simple models, is the coupled 

rotator Slowly Relaxing Local Structures model. However, when slow 

motions about the glycosidic linkage, and of the hydroxymethyl group, 

are studied in small oligosaccharides, which are the systems of interest 

to us, distinguishing individual rotators is problematic. We developed 

a new stochastic model which is applicable to flexible rotators with 

one internal degree of freedom. This approach combines stochastic 

mesoscopic treatment of the comprehensive diffusion operator in the 

presence of a local potential, hydrodynamic calculation of the global 

diffusion, and atomistic DFT calculations of the local potential (for our 

oligosaccharides rather large basis sets can be used). The only free 

parameter is the bead radius in the hydrodynamic calculation. This 

model was tested for the disaccharide β-D-glucopyranoside (1→6) 

α-D-(6- 13 C) mannose-OMe dissolved in a DMSO-d 6 /D 2 O cryosolvent. 

NMR relaxation parameters were measured for the linkage 13 CH 2 probe. 

The experimental data acquired include 13 C T 1 , T 2 and 13 C-{ 1 H} NOE, 

as well as longitudinal and transverse dipole-dipole cross-correlated 

relaxation rates, acquired over a broad temperature range. While 

previous attempts using simple models could not fit these data, our new 

model reproduced them very well. 

Re20 

Investigating complex materials with relaxation of fluorinated 

gases 

Lounila, Juhani; Tervonen, Henri; Jokisaari, Jukka 

University of Oulu, Department of Physical Sciences, Oulu, Finland 

Useful information about the structures of complex materials (e.g., 

porous media) may be extracted from several NMR parameters. For 

example, the spin-lattice relaxation times (T 1 ) of fluorine nuclei in inert 

gases such as CF 4 and C 2 F 6 are known to increase considerably when 

the gases are confined to small pores [1]. Such behaviour is expected 

for collisionally interrupted intramolecular interactions in the limit of 

extreme narrowing. 

As a matter of fact, the dominant relaxation mechanism for these gases 

is the modulation of the spin-rotation interaction by molecular collisions. 

In the bulk gas, the correlation time of molecular angular momentum is 

determined only by the molecule-molecule collisions, and therefore in 

the fast correlation limit T 1 is proportional to the density of the gas. In 

porous materials, the collision frequency is increased by the presence of 

molecule-wall collisions. Hence, the increase of the relaxation time may 

be attributed to the increase of the collision frequency [1]. 

A method for measuring the surface/volume ratio of porous materials 

by measuring how T 1 of CF 4 gas changes with confinement has recently 

been introduced [2]. The applicability of the method was demonstrated 

by analyzing samples of fumed silica (SiO 2 ). This material is an ultrafine 

powder with a high air content and very little particle-particle contact, 

and its surface/volume ratio can be changed by compression. 

However, in most practical applications the studied material is granular. 

Then the analysis is complicated by the presence of voids where the 

gas molecules can reside. We have searched for a generalization of the 

method to macroscopically inhomogeneous materials by studying the 

19 F spin-lattice relaxation time of sulfur hexafluoride gas (SF6 ) adsorbed 

in different types of granular porous materials. 

[1] M. J. Lizak, M. S. Conradi, and C. G. Fry, J. Magn. Reson., 95, 548 

(1991). 

[2] D. O. Kuethe, R. Montano, and T. Pietrass, J. Magn. Reson., 186, 

243 (2007). 


Re21 

Characterization of unsaturated porous media by high-field and 

low-field NMR relaxometry 

Stingaciu, Laura-Roxana 1 ; Pohlmeier, Andreas 1 ; Weihermüller, Lutz 1 ; 

Vereecken, Harry 1 ; Blümler, Peter 2 ; van Dusschoten, Dagmar 2 ; Stapf, 

Siegfried 3 

1 Forschungszentrum Jülich, Agrosphere Institute, ICG-4, Jülich, 

Germany; 2 Forschungszentrum Jülich, ICG-3, Jülich, Germany; 

3 University of Technology, Dept. of Technical Physics II, Ilmenau, 

Germany 

A comparison study of Nuclear Magnetic Resonance Relaxometry 

at high and low magnetic field (7 T and 0.1 T) has been initiated for 

investigating the influence of the magnetic field strength; variable 

clay content and different degrees of saturation on the relaxometric 

properties of four ideal porous media. The samples consisted of 

medium sand with increasing fractions of kaolin clay ranging from 0 to 

15%. Six different volumetric water contents between saturation and 

θ = 0.05 were used. Changes in water content of the samples were 

achieved by slow evaporation. T2 relaxation curves were monitored by 

the Carr-Purcell-Meiboom-Gill sequence (CPMG) and further analyzed 

by inverse Laplace transformation yielding T2 distribution functions. 

Sand shows a slight continuous shift with decreasing water content of 

a bimodal distribution function of T2 to faster relaxation at high and low 

magnetic field. Sand clay mixtures show broad, bimodal distribution 

functions for both magnetic field intensities which shift slightly with 

decreasing water content. The relaxivity in these systems is surface 

dominated, as the weak and comparable dependence of 1/T2 on TE 

at both field strengths show. Signal amplitude behaviour with variation 

of saturation degree was also monitored. An expected proportionality 

of the total signal amplitude with water content was observed for all 

samples at 0.1 T whereas at 7 T deviations occurred for samples with a 

clay content higher than 5%, which are assigned to loss of signal in the 

first echo periods. 

Re22 

Gadolinium-loaded gold nanoparticles as contrast 

agents for MRI 

Warsi, Muhammad; Chechik, Victor 

The University of York, Chemistry, York, United Kingdom 

Magnetic resonance imaging (MRI) is a powerful diagnostic technique 

used in modern biomedical research. MRI is based on nuclear magnetic 

resonance (NMR). It has many advantages over other techniques 

as it is non-invasive and has excellent spatial resolution. The use 

of contrast agents (CAs) further enhanced its success by changing 

the signal intensities. The contrast agents (CAs), mostly gadolinium 

based, affect the intensities of proton NMR signals by altering the 

relaxation rate of (water) protons in the body. The maximum relaxivity 

can be achieved by slowing down the tumbling of contrast agent 

molecules and attaching several gadolinium ions per molecule. To 

get maximum relaxivity, gold nanoparticles (AuNPs) stabilized by new 

DTPA (diethylenetriaminepentaacetic acid) based ligand were prepared 

and then gadolinium ions were loaded. The spin lattice relaxivity (R1) 

of Gd-loaded-AuNPs was measured and found 27% higher than the 

commercially available MRI contrast agent i.e. Gd-DTPA. Further 

increase (~70%) in R1 of Gd-loaded-AuNPs was found by forming 

polyethyleneimine (PEI) layers around AuNPs (that reduced the tumbling 

of AuNPs). The rotational correlational time (τcorr) of AuNPs was 

measured by vanadyl (VO2+) labelling using EPR measurements. EPR 

measurements were found compatible with NMR measurements. A 

recognition vector (biotin terminated thiol) was attached on AuNPs; the 

efficiency of molecular recognition was shown in a proof-of-principle 

experiment. 


Re23 

Simulation of NMR relaxation induced by superparamagnetic 

particles in solution 

Vuong, Quoc Lam 1 ; Roch, Alain 2 ; Gillis, Pierre 1 ; Gossuin, Yves 1 

1 University of Mons, Biological Physics Department, Mons, Belgium; 

2 University of Mons, Mons, Belgium 

The knowledge of Nuclear Magnetic Resonance (NMR) relaxation times 

is of great importance in several MR (Magnetic Resonance) applications. 

For example, in MR Imaging, contrast agents are often used to modify T1 

and T2 of body water: This results in an improved MR image contrast. 

Superparamagnetic particles constitute one important class of contrast 

agents: The dipolar interaction of their large magnetic moment with 

proton spin considerably influences proton relaxation. Several theories 

have attempted to predict them in homogeneous colloidal solution in 

the last decade [3,5]: Classical Redfield theories coupled with Néel 

relaxation [1] is used. 

However, these theories do not consider three important characteristics 

of the magnetic particles that can not be treated analytically. (1) The real 

dynamic of electron spins (2) Particles with large radius that are not in 

the Redfield conditions (3) Clustering that appears when particles are 

injected in vivo. 

In this work, we study the influence of these parameters in T1 and T2 

using simulations. Dynamic of electron spins is obtained by simulating 

the Brown Equation [2]. This allows quantifying Néel relaxation time and 

Larmor frequency variations that influence relaxation times. Monte Carlo 

simulations are used to study the transverse relaxation induced by large 

particles and by clustered particles as previously described [4]. These 

simulations give T2 at high fields. 

[1] L. Néel, Ann. Geophys. (C.N.R.S.), 5,99-136, 1949. 

[2] W.F. Brown, Phys Rev 130, 5, 1677-1686, 1963. 

[3] A. Roch, R.N. Muller, P. Gillis, Journal of Chemical Physics 110, 11, 

5403-5411, 1999. 

[4] P. Gillis, F. Moiny, R.A. Brooks, Magn Reson Med, 47, 257-263, 2002. 

[5] Y. Gossuin, P. Gillis, A. Hocq, Q.L. Vuong and A. Roch, WIREs 

Nanomed Nanobiotechnol 1, 299-310, 2009. 

Re24 

Long Lived Coherences in High Field NMR 

Sarkar, Riddhiman; Ahuja, Puneet; Vasos, Paul R.; Bodenhausen, 

Geoffrey 

Ecole Polytechnique Fédérale de Lausanne, Chemistry and Chemical 

Engineering, Lausanne, Switzerland 

Long Lived States 1,2 (LLS) do not evolve under coherent interactions. 

They often have much longer lifetimes than spin lattice relaxation time 

constants. On the contrary, for scalar coupled two spin ½ systems, 

coherent superpositions of singlet and the central triplet states 3 , which 

can be called Long Lived Coherences (LLC), oscillate with the frequency 

of the scalar coupling and have lifetimes much longer than conventional 

transverse relaxation time constants (T LLS >T LLC >T 2 ). These oscillations 

can be monitored in the manner of 2D spectroscopy leading to very 

narrow lines with half-widths at half-height of 60 mHz which is beyond 

the scope of conventional NMR spectroscopy. This is not only true for 

small molecules but can be demonstrated in the highly mobile Glycine - 

76 residue in ubiquitin. 

References 

[1] M. Carravetta and M.H. Levitt, 

J. Am. Chem. Soc. (2004), 126, 6228-6229 

[2] R. Sarkar, P. R. Vasos, and G. Bodenhausen, 

J. Am. Chem. Soc. (2007), 129, 328-334 

[3] G. Pileio, oral presentation, EUROMAR 2008 

ABSTRACTS POSTER


Re25 

Damped quantum rotation in NMR spectra. Numerical 

simulations for the cyclobutadienyl dication ring 

Ratajczyk, Tomasz; Szymanski, Slawomir 

Institute of Organic Chemistry PAS, Warszawa, Poland 

The damped quantum rotation (DQR) theory describes manifestations 

in NMR spectra of the coherent and stochastic dynamics of hindered 

molecular rotors composed of indistinguishable nuclei. Formulated 

first for methyl-type rotors, it was recently generalized to N-fold 

systems such as the aromatic hydrocarbons’ rings. In the familiar 

Alexander-Binsch (AB) line-shape theory, the relevant stochastic 

processes are pictured as classical random jumps of the rotor between 

its N equilibrium orientations. The DQR model employs a quantum 

mechanical approach based on the Pauli principle correlating the spatial 

and nuclear-spin degrees of freedom of the rotor. Here, the processes 

evidenced in NMR spectra are damped oscillations of certain quantum 

coherences between its spin-space correlated torsional states. These 

are specific, long-lived coherences resistant to destruction under impact 

of the condensed environment. For N > 2, the relevant damping-rate 

constants outnumber the classical rate constants of direct jumps over 

one, two, etc potential maxima. Nevertheless, when the magnitudes 

of the former fit appropriate “classical” patterns, the jump picture is 

recovered. For the methyl groups in both solid- and liquid-phase NMR, 

the DQR approach has already been confirmed experimentally. A part 

of the DQR approach is a theoretical model of temperature effects 

on the quantum rate constants. For a given system it enables one to 

assess the extent of its possible departure from the classical limit. In 

the present contribution, perspectives of detecting the DQR effects in 

NMR spectra of solids containing a hindered cycobutadienyl dication 

are considered. Numerical calculations of the DQR rate constants for 

this model system, which is the simplest aromatic ring of N>3, are 

performed. On this basis, theoretical single-crystal spectra of the ring 

protons are simulated. Attemtps at fitting these spectra with the AB 

equation reveal that even for such a “sizable” molecular rotor there are 

real possibilities of detecting the DQR effects in NMR spectra. 

Re26 

Measuring transverse relaxation rates in J-coupled systems 

Aeby, Nicolas; Baishya, Bikash; Segawa, Takuya; Bodenhausen, Geoffrey 

Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland 

Measuring transverse relaxation rates R 2 in spin systems with 

homonuclear J-couplings is a challenge: modulations due to the 

J-couplings tend to mask the exponential echo decays. A recent 

method to extract R 2 ’s from monoexponential decays uses Carr-Purcell- 

Meiboom-Gill (CPMG) -trains with moderate RF-field amplitudes (ν 1 

comparable to the difference in offsets), while avoiding so-called 

“recoupling” conditions (1,2) . For J-coupled multi-spin systems, 

monoexponential decays can be obtained by adequate positioning of 

the carrier frequency. This is demonstrated for 13 C in uniformly 13 C- and 

15 N-labelled Alanine and for 1 H in Cyclosporin A. 

References 

[1] K. Gopalakrishnan, N. Aeby and G. Bodenhausen, Chem. Phys. 

Chem. (2007), 8, 1791-1802. 

[2] N. Aeby and G. Bodenhausen, Chem. Phys. Lett. (2008), 463, 418- 

421. 

92 

Re27 

Inclusion complexes of a cryptophane – A derivative and 

cryptophane – C with 13 CHCl 3 : An NMR study of exchange 

kinetics and relaxation 

Takacs, Zoltan 1 ; Soltesova, Maria 2 ; Brotin, Thierry 3 ; Dutasta, Jean- 

Pierre 3 ; Kowalewski, Jozef 1 

1 Stockholm University, D. of Physical, Inorganic and Structural 

Chemistry, Stockholm, Sweden; 2 Charles University, Faculty of 

Mathematics and Physics, D. of Low Temperature physics, Prague, 

Czech Republic; 3 Ecole Normale Superieure de Lyon, Laboratoire de 

Chimie, Lyon, France 

Cryptophanes play an important role in supramolecular chemistry. They 

are composed of two cup – shaped cyclotribenzylene units which are 

bound together by three aliphatic linkages. This structure provides a 

nearly spherical three-dimensional cavity which is able to bind small 

organic molecules such as chloroform or dichloromethane. The special 

molecular recognition properties of these compounds make it possible 

to use them as storage of small molecules. The present study focuses 

on the complexes formed between two cryptophanes, cryptophane- A 

with methoxy groups replaced by butoxy units and cryptophane – C, 

and carbon-13 labeled chloroform. The two cyclotribenzylene units in 

the former cryptophane are equivalent, while they differ in cryptophane 

– C. There, one of the cyclotribenzylene units carries the methoxy 

groups on the phenyl rings, while the substituents are absent in the 

other end of the molecule. This property makes it possible to analyze 

the orientation of chloroform inside the cavity by the 2D NOESY and 

ROESY experiments. The kinetics of the complex formation was studied 

by using one-dimensional magnetization transfer, 1D – EXSY type of 

experiments. From the measurements of the rate constants at different 

temperatures, the activation energies were estimated using the simple 

Arrhenius plot. 13 C relaxation measurements (T 1 , NOE) were performed 

at 9.4 and 14.1 T for both the hosts and the guest. In the interpretation 

of the guest relaxation data, the exchange kinetics needs to be taken 

into consideration. From the 13 C relaxation properties, it is possible to 

investigate the motion of the guest molecule inside the host cavity. 

Re28 

Intermolecular dipole-dipole cross relaxation. Small molecular 

solute-solvent systems 

Nordstierna, Lars 

Chalmers University of Technology, Department of Chemical and 

Biological Engineering, Göteborg, Sweden 

Solute-solvent interactions have been studied by { 1 H} 1 H and { 1 H} 19 F 

NMR spectroscopy. Different solutions of tri- and tetrahalomethanes 

were investigated with the purpose to obtain model systems with simple 

non-charged molecules. Other desirable conditions for the molecular 

structure were spherical geometry and solely one 1 H or 19 F nucleus 

per molecule. In the experiments, intermolecular dipole-dipole crossrelaxation 

rates between specific nuclei were recorded. The relaxation 

was examined for two types of systems. First, the homonuclear cross 

relaxation between 1 H of the CHI 3 solute and the CHBr 3 solvent. Second, 

the heteronuclear cross relaxation between 19 F of the CFBr 3 solute and 

1 H nuclei of the CHBr3 solvent. The experiments were performed at 

several magnetic field strengths ranging from 4.70 up to 18.81 Tesla. 

The results reveal a significant frequency dependence of the cross 

relaxation even for small and fast tumbling molecules. The decrease 

of the cross-relaxation rate with respect to the NMR frequency agrees 

with earlier results where ionic surfactant interactions with water were 

studied. 1,2 The findings clearly validate the lack of extreme narrowing 

for nuclear spin relaxation by diffusionally-modulated intermolecular 

interactions. The magnetization transfer between solvent molecules 

and the solute is not solely controlled by fast local motions but also 


significantly contributed by long-range translational dynamics. 

The experimental data were evaluated within the framework of models 3 

for diffusionally-modulated relaxation. Supplementary longitudinal 

relaxation and self-diffusion experiments were recorded in order to aid 

the evaluation. The model provides information about the degree of 

dynamic retardation of the solvent molecules that are in contact with the 

solute. Given sufficiently field-dependent experimental data, the number 

of layers with retarded solvent molecules can be calculated. 

References 

(1) Nordstierna, L.; Yushmanov, P. V.; Furó, I. J.Chem.Phys 125:074704 

(2006) 

(2) Nordstierna, L.; Yushmanov, P. V.; Furó, I. J.Phys.Chem.B 110:25775 

(2006) 

(3) Halle, B. J.Chem.Phys. 119:12372 (2003) 

Small Molecules 


Sm10 

Improved properties of high molecular weight polyglutamates 

as chiral orienting media for organic compounds 

Marx, Andreas; Thiele, Christina 


Germany 

The determination of relative configurations in organic compounds 

by NMR does not always lead to unambiguous results. The 

recently reintroduced residual dipolar couplings (RDCs) [1] provide 

complementary information to these conventional NMR restraints and 

can lift ambiguities [2]. 

In order to be able to observe RDCs the solute of interest must be 

oriented with respect to the magnetic field. Orienting media for organic 

compounds are still scarce, with the situation being even worse, if chiral 

media are considered. These could potentially allow the determination 

of absolute configurations in the future. So far only homopolypeptides 

have been applied successfully as chiral orienting media for organic 

compounds. The degree of orientation of commercially available 

homopolypeptides, however, tends to be too large, which makes a 

reliable determination of RDCs difficult. 

We have synthesized a series of polyglutamates and could show 

that high molecular weight PBLGs induce a much lower degree of 

orientation. These much improved orienting properties make the reliable 

extraction of RDCs possible. We observed that although the degree of 

orientation is changed significantly, enantiodiscrimination is independent 

of the molecular weight of PBLG [3]. 

Therefore we wanted to get a deeper insight into factors responsible 

for enantiodiscrimination and had a closer look on how different 

the diastereomorphous orientations for one example compound 

(isopinocampheol) are in these helically chiral orienting media. 

Therefore, the change in solute orientation between poly-γ-benzyl-Lglutamate 

(PBLG) and its enantiomer poly-γ-benzyl-D-glutamate (PBDG) 

was investigated in detail [4]. 

References: [1] Reviews (organic compounds): C.M. Thiele, Conc. Magn. 

Reson. 2007, 30A, 65–80; C. M. Thiele, Residual Dipolar Couplings 

(RDCs) in Organic Structure Determination, Eur. J. Org. Chem. 2008, 

5673-5685. 

[2] C.M. Thiele, A. Marx, R. Berger, J. Fischer, M. Biel, A. Giannis, Angew. 

Chem. Int. Ed. 2006, 45, 4455–4460. 

[3] A. Marx, C.M. Thiele, Chem. Eur. J. 2009, 15, 254–260. 

[4] A. Marx, V. Schmidts, C.M. Thiele, submitted 2009. 

Sm11 

Determination of the conformation of the Key-Intermediate of 

the Pd-catalyzed allylic substitution from RDCs 

Böttcher, Benjamin; Schmidts, Volker; Thiele, Christina 


Germany 

Residual Dipolar Couplings (RDCs) are becoming increasingly important 

in the determination of configuration and conformation of small 

organic compounds [1]. Methods are being developed to also include 

conformational flexibility into these analyses [2, 3]. So far, however, 

RDCs have not been applied to gain insight into reaction mechanisms. 

We want to report on the determination of the conformation of the 

reactive intermediate of the enantioselective Pd-catalyzed allylic 

substitution, which could not be determined based on conventional 

NMR restraints. 

ABSTRACTS POSTER


We therefore oriented the very sensitive complex in high molecular 

weight PBLG [4] to obtain RDCs of suitable size. After measuring 

several 1 D C-H we fitted these data to several possible diastereomorphous 

representations of conformers. This resulted in a much better fit for one 

conformer than for the others. As the fit, however, was not excellent, we 

examined each fragment of the complex individually and determined 

their orientation with respect to each other using local orienting tensors 

[3, 5]. 

References: 




Angew. Chem. Int. Ed. 2006, 45, 4455-4460; A. Schütz, J. Junker, 

A. Leonov, O. F. Lange, T. F. Molinski, C. Griesinger, J. Am. Chem. Soc. 

2007, 129, 15114-15115. 

[3] C. M. Thiele, A. Maliniak, B. Stevensson, unpublished results. 

[4] A. Marx, C. M. Thiele, Chem. Eur. J. 2009, 15, 254-260. 

[5] B. Böttcher, V. Schmidts, J. Raskatov, C. M. Thiele, unpublished 

results; B.Ulriksen (nee Laursen), J. Raskatov, H. Steinhagen, A. Geiser, 

B. Wiese, C. Mürmann, B. Böttcher, V. Schmidts, C. M. Thiele, M. 

Reggelin, G. Helmchen, unpublished results. 

Sm12 

Conformational analysis of some carbasugars by NMR 

spectroscopy 

Säwén, Elin; Roslund, Mattias; Cumpstey, Ian; Widmalm, Göran 

Stockholm University, Organic chemistry, Stockholm, Sweden 

Idose and iduronic acid are unusual among hexoses in that their 

pyranose forms do not exist exclusively in a single chair conformation 

but are rather flexible, having several low energy conformations. 

Iduronic acid is a component of various biologically active glycans, and 

the ring flexibility may be the key to the strong binding of these glycans 

to their receptors. 

Carbasugars are carbohydrate analogues in which the endocyclic ring 

oxygen has been replaced by a methylene group. This means that the 

acetal linkage of a glycoside is formally transformed into an ether for 

a carbasugar, and hence becomes hydrolytically stable. Two different 

carbaiduronic acid derivatives were therefore synthesized. To assign the 

ring conformations in solution of the carbaiduronic acids different NMR 

techniques as well as NMR spin simulation were used to extract 3 J H,H , 

2 JC,H and 3 J C,H and proton – proton distances. 

3 JH,H coupling constants were extracted from 1D 1 H NMR spectra 

and refined by spin simulations with the PERCH NMR software. 

Heteronuclear long-range 1 H, 13 C coupling constants are very important 

parameters in the determination of conformations. Three different 2D 

NMR techniques have been used to investigate the magnitudes and/ 

or signs of the coupling constants, namely, J-HMBC, HSQC-HECADE 

and the IPAP-hadamard-HSQC-TOCSY experiment with zero quantum 

suppression. To determine proton – proton distances 1D NOESY and 1D 

T-ROESY experiments were used. 

94 

Sm13 

NMR investigation of reaction 1,4-dihydroxynaphtholine 

adamantilation 

Peterson, I.V. 1 ; Sokolenko, W.A. 1 ; Svirskaya, N.M. 1 ; Kondrsenko, A.A. 2 ; 

Rubailo, A.I. 2 

1 Institute of Chemistry and Chemical Technology, RAS (Sib. branch), 

Krasnoyarsk, Russian Federation; 2 Siberian Federal University, 

Krasnoyarsk, Russian Federation 

Adamantane, 1,4-naphthoquinone, 1,4-dihydroxynaphtholine and they 

derivatives possess wide spectrum of pharmacological activity [1-3]. 

Adamantilation reaction of 1,4-dihydroxynaphtholine was investigated. 

The assignment of signal of protons and carbons atoms for 

2-(1-adamantile)-2,3-dihydronaphthoquinone-1,4 (I) and 

2-(1-adamantile)-naphthoquinone-1,4 (II) was carried out using 1H and 

13C spectroscopy. 

To attribute protons H , 2 IH and 3 IIH in compound I the following torsion 

3 

angels were measured: φ (H -C -C - 1 2 2 3 IH )=67,179°; φ (H -C -C - 

3 2 2 2 3 

IIH3 )=174,123° and angel IH -C - 3 3 IIH =107,108°. 

3 

Vicinal and geminal constant of spin-spin interaction were calculated 

using Karplus equation [4]: 3J =3.0, (ax,eq) 3J =6,6 and (ax,ax) 2J= -17,01 Hz. 

1H (δ, ppm): H2 =2.74, IH =3.22 and 3 IIH =3,01. 3 13C (δ, ppm): C =59.42, 

2 

C =38,87. 

3 

For compound II 1H (δ, ppm): H =7.41. 3 13C (δ, ppm): C =158.55, 

2 

C =134,83. 

3 

References 

[1] Pearson M., Jenshky B. J. Org. Chem., 1978. Vol.43, p. 4617-4622. 

[2] Oxford, J.S., Pharmaceutical Therapy, 1981. Vol. 11, p. 181-262. 

[3] Laatsch. H., Liebigs Ann. Chem. 1980. p.140-157. 

[4] Keisuke Imai, Magnetic Resonance in Chemistry, 1989. Vol.28, p. 

668-674. 

Sm14 

29 13 Si- C Couplings along the Si-O-C-C-C chain in aromatic and 

aliphatic compounds 

Schraml, Jan1 ; Blechta, Vratislav1 ; Sykora, Jan1 ; Kurfurst, Milan1 ; 

Sychrovsky, Vladimir2 1Institute of Chemical Process Fundamentals of the ASCR, v. v. 

i., Prague, Czech Republic; 2Institute of Organic Chemistry and 

Biochemistry of the ASCR, v. v. i., Prague, Czech Republic 

Several methods were developed for determination of the signs and 

magnitudes of small indirect J-couplings between 29 Si and 13 C nuclei 

separated by two, three, and four bonds. The methods took advantage 

of the specific features of 29 Si NMR, namely long T 2 relaxation times 

and large spectral dispersion; some of the methods overcome absence 

of protons on quaternary carbons or nil 29 Si- 1 H couplings with protons 

separated from the 29 Si atom by more than three bonds. Using these 

methods series of aromatic and aliphatic compounds were studied 

and empirical rules for relative magnitudes of the J-coupling constants 

were established to aid the assignments based on 29 Si- 13 C correlation 

experiments. The experimental findings were supported by DFT 

calculations. 

Acknowledgement: This work was supported by the Grant Agency of 

the Academy of Sciences of the CR (grant Nos. IAA400720706 and 

IAA400550701). 


Sm15 

NMR studies on Cu(III) intermediates in S N -reactions of 

sterically hindered organocuprates 

Neumeier, Maria; Gärtner, Tobias; Gschwind, Ruth M. 

Universität Regensburg, Organic Chemistry, Regensburg, Germany 

Regio- and diasteroselective C-C bond formation is one of the most 

important tools in organic synthesis. Therefore, organocopper reagents 

are frequently used in cross coupling reactions with alkyl halides or 

addition reactions with Michael acceptors. The key intermediate in 

these copper mediated reactions has long been believed to be a Cu(III) 

intermediate due to theoretical calculations supporting this hypothesis 

[1]. However, the experimental evidence for these elusive Cu(III) 

intermediates had been missing for a long time. 

Since the independent NMR detection of the first stabilised Cu(III) 

intermediates by our group and that of Bertz and Ogle [2,3], we focus 

on the influence of sterically demanding residues. 

With detailed NMR spectroscopic investigations it is possible to study 

the effects of steric hindrance on the reactivity of organocuprates which 

is shown in this presentation. Furthermore, the influence of electron 

donating ligands on the stability of the decisive Cu(III) intermediate in 

solution is discussed, based on clear NMR spectroscopic results. 

[1] E. Nakamura, S. Mori (2000) Angew. Chem. Int. Ed. 39, 3750-3771 

[2] S. H. Bertz, S. Cope, M. Murphy, C. A. Ogle, B. J. Taylor (2007) J. Am. 

Chem. Soc. 129, 7208-9 

[3] T. Gärtner, W. Henze, R. M. Gschwind (2007) J. Am. Chem. Soc. 129, 

11362-3 

Sm16 

Pd-catalyzed alkylation and dimerization of olefins by timeresolved 

NMR spectroscopy 

John, Michael; Sachse, Anna; Meyer, Franc 

Georg-August-Universität, Institut für Anorganische Chemie, Göttingen, 

Germany 

Dinuclear Pd complexes with a defined metal-metal distance bear 

high potential as catalysts in olefin oligo- and polymerization. Here we 

present a novel tetranuclear complex 1 bearing a Pd-CH 2 -Pd bridge 

that could be generated from a dinuclear Pd-Cl species A by reaction 

with SnMe 4 . The mechanism of this reaction was studied using various 

NMR techniques (including NOESY and diffusion NMR) and found to 

proceed via two Pd-Me intermediates that combine under elimination 

of methane. We further studied the reactivity of 1 with olefins by timeresolved 

NMR spectroscopy and found that 1 is able to both alkylate 

olefins and catalyze their dimerization while A is reformed. The details 

of this reactivity were investigated by analyzing the products of the 

reaction of 1 with [ 1 H, 12 C], [ 1 H, 13 C] and [ 2 H, 12 C]-labeled ethylene as well 

as propylene. 

Sm17 

Solution Structure of a Central Fragment of Tumor Antigen 

Le a Le x 

Svensson, Mona 1 ; Jackson, Trudy A 2 ; Wang, An 2 ; Widmalm, Göran 1 ; 

Auzanneau, France-Isabelle 2 

1 Stockholm University, Organic Chemistry, Stockholm, Sweden; 

2 University of Guelph, Department of Chemistry, Ontario, Canada 

The trisaccharide β-D-GlcpNAc-(1→3)-β-D-Galp-(1→4)-β-D- 

GlcpNAc-OMe is a central fragment of the Le a Le x hexasaccharide [1] , 

which is commonly expressed by squamous lung carcinoma cells. 

Using the Le a Le x hexasaccharide as an antigen would likely induce an 

autoimmune reaction since the terminal non-reducing Le a trisaccharide 

is commonly expressed by healthy tissue. However, some monoclonal 

antibodies raised against Le a Le x have been demonstrated to be 

specific for internal epitopes of the hexasaccharide while showing no 

cross-reactivity with Le a . It is therefore of interest to identify and fully 

characterize such epitopes, thereby permitting the development of 

the necessary targeted vaccines. Here we interpret conformational 

information obtained for the above trisaccharide, using the 2D 

J-HMBC [2] experiment which gives information on trans-glycosidic 3 J C,H 

and 1D 1 H, 1 H-NOESY experiments. 

[1] Jackson, T. A.; Robertson, V.; Imberty, A.; Auzanneau, F. -. Bioorganic 

and Medicinal Chemistry 2009, 17, 1514-1526. 

[2] Meissner, A.; Sørensen, O. W. Magnetic Resonance in Chemistry 

2001, 39, 49-52. 


Sm18 

Determination of conformer populations from residual dipolar 

couplings 

Schmidts, Volker 1 ; Thiele, Christina 1 ; Böttcher, Benjamin 1 ; Louzao, Iria 2 ; 

Berger, Robert 3 ; Maliniak, Arnold 4 ; Stevensson, Baltzar 4 

1 Technische Universität Darmstadt, Clemens Schöpf Institut, Darmstadt, 

Germany; 2 Universidad de Santiago de Compostela, Departamento 

de Quimica Organica, Santiago de Compostela, Spain; 3 Universität 

Frankfurt, Frankfurt Institute for Advanced Studies, Frankfurt, Germany; 

4 Stockholm University, Division of Physical Chemistry Arrhenius Lab, 


For the determination of the three dimensional structure of organic 

molecules by NMR spectroscopy usually 3 J couplings, NOEs and crosscorrelated 

relaxation data are used. Recently, it has been shown that 

residual dipolar couplings (RDCs) can yield complementary information 

to these NMR observables. [1] 

In the case of flexible molecules only averaged observables can 

be extracted due to fast (on the timescale of NMR interactions) 

interconversion of conformers. The unambiguous determination of the 

configuration and conformation from averaged J couplings and NOEs 

is often impossible as is the case for the α-Methylene-γ-butyrolactone 

investigated here.[2, 3] It can however, be resolved when using RDCs. 

It can exists in two conformational states per diastereoisomer. We have 

investigated several routes for the treatment of the conformational 

equilibrium. Depending on the dynamics of the system, two situations 

may be identified: i) a common order tensor for all conformers is defined 

or ii) one tensor for each conformer is required.[4] The conformer 

populations extracted using both approaches are in excellent agreement 

with those obtained from quantum chemical calculations and computed 

3 J couplings. This also indicates that the conformational equilibrium is 

not significantly shifted by the orienting medium. 

References: 




Angew. Chem. Int. Ed. 2006, 45, 4455-4460. 

[3] C. M. Thiele, V. Schmidts , B. Böttcher, I. Louzao, R. Berger, A. 

Maliniak, B. Stevensson, to be published. 

[4] E. E. Burnell, C. A. de Lange, J. Magn. Reson. 1980, 39, 461-480. 

ABSTRACTS POSTER


Sm19 

The self-assembly behavior of small organic molecules in water 

using NMR and light scattering techniques 

Odeh, Fadwa 1 ; Al-Bawab, Abeer 1 ; Li, Yuzhuo 2 

1 University of Jordan, Amman, Jordan; 2 Clarkson University, Potsdam, 

United States 

The self-assembly behavior of 1H-benzotriazole (BTA) and 

benzotriazole-1-methanol (BTAOH) in aqueous solutions below their 

solubility limit has been revealed for the first time using NMR and light 

scattering techniques. Relaxation time, diffusion and chemical shift NMR 

techniques in addition to various light scattering techniques were used 

to study the aqueous behavior of BTA and BTAOH. These studies have 

revealed the self assembly of these molecules in water. Results show 

that BTA molecules tend to aggregate in water to form nanoparticles 

with radius in the range of 5 nm. The critical aggregation concentration 

(CAC) is estimated based on NMR data to be ~16-20 mM. Such 

a critical aggregation concentration is comparable with the typical 

critical micelle concentration (CMC) for surfactants that have moderate 

aqueous solubility. The self-assembly behavior of BTA and BTAOH 

may not be limited to benzotriazoles. It might be generally true for all 

poorly water soluble species to aggregate at concentrations below their 

solubility limits. 

Sm20 

Conformational diversity in thiourea based bifunctional 

organocatalysis 

Király, Péter 1 ; Vakulya, Benedek 2 ; Varga, Szilárd 2 ; Soós, Tibor 2 ; Tárkányi, 

Gábor 1 

1 Chemical Research Center, NMR Spectroscopy, Budapest, Hungary; 

2 Chemical Research Center, Synthetic Organic Chemistry, Budapest, 

Hungary 

We have recently discovered that bifunctionality, an essential factor in 

organocatalysis is also an inherent source of intermolecular interactions 

for the thiourea modified members of the cinchona family [1]. As part of 

our ongoing research in understanding the structure-function as well as 

the kinetics-yield relation for a variety of catalytic systems we aimed to 

investigate the conformational dynamics of a set of structurally similar 

catalysts. Low temperature solution NMR spectroscopy has become 

a valuable method [1] in exploring the nature of thiourea CSNH--- 

:N hydrogen bonding within these systems. Some members of the 

catalyst family form self-assembled dimers via intermolecular hydrogen 

bonding whereas others prefer intramolecular hydrogen bonding. This 

interplay between inter- and intramolecular interactions has turned 

our attention towards the structural as well as stereochemical reasons 

of the formation of non-hydrogen bonded “unnatural” conformers 

regarded as catalytically hot species. In the present work we describe 

the conformational diversity of six catalysts on the basis of field 

dependent exchange NMR and H–H spatial correlations from low 

temperature (< –40°C) NOESY and ROESY experiments. As we explored 

the thermodynamics of the transient dimer formation we discovered 

the possibility of higher order molecular organization an important 

phenomenon when fine tuning the reaction conditions under high 

catalyst loading. A particularly interesting aspect of this work is the 

discovery of monomer-dimer interactions. 

References 

[1] Tárkányi et. al. Chem. Eur. J. 2008, 14, 6078–6086. 

Acknowledgements - The support of the Hungarian GVOP-3.2.1.-2004- 

04-0210/3.0 project and OTKA K-69086 are gratefully acknowledged. 

96 

Sm21 

Relative configuration of a conformationally flexible molecule in 

solution determined by RDC- and NOE-enhanced NMR 

Sun, Han 1 ; Reinscheid, Uwe 1 ; d’Auvergne, Edward 1 ; Oliveira Rocha, 

Rafael 2 ; Kleber Z.Andrade, Carlos 2 ; Carlos Dias, Luiz 3 ; Griesinger, 

Christian 1 

1 Max-Planck-Intitute for biophysical chemistry, NMR-based structural 

biology, Abt.10300, Göttingen, Germany; 2 Universidade de Brasilia, 

Brasilia, Brazil; 3 Universidade Estadual de Campinas, Campinas, Brazil 

Determination of relative configuration of natural products or organic 

molecules in general is often carried out by X-ray diffraction [1] for 

compounds that can be crystallized, by chemical synthesis or by 

solution-state NMR spectroscopy. Although conventional scalar coupling 

and NOE analysis has been used successfully for discrimination of 

inflexible diastereomers, application to flexible molecules has not yet 

been possible. 

In this study, the stereochemical elucidation of a very flexible Evan’s 

product is presented. The two neighboring, unknown stereocenters 

together with several rotatable bonds in the molecule complicate the 

problem. As a consequence, the number of possible diastereomers can 

only be reduced from four to two by standard J-coupling analysis. By 

measuring residual dipolar couplings (RDCs) [2] in the well established 

alignment media PH-gel [3] and using rotating-frame NOE (ROEs) 

restraints, the two diastereomers can be discriminated accurately. This 

method utilises numerous ensembles of discrete conformations derived 

from MD trajectories and answers the stereochemical question using 

the rotating-frame NOEs (ROEs) and RDCs to differentiate between 

configurations. 

[1] Bijvoet, J. M., Peerdeman, A. F. & Vanbommel, A. J. (1951). Nature 

168, 271-272. 

[2] A. Bax, Protein Science 2003, 12, 1-16; J. H. Prestegard, C. M. 

Bougault, A. I. Kishore, (2004). Chem. Rev., 104, 3519-3540; C. 

Griesinger, J. Meiler, W. Peti, (2003). Biol. Magn. Res. 20, 163-229. 

[3] P. Haberz, J. Farjon, C. Griesinger, (2005). Angew. Chem. Int. Ed., 44, 

427-429. 

Sm22 

Applications of 1D and 2D INADEQUATE NMR spectroscopy for 

background signal suppression in studying the reactions of the 

vesicant sulfur mustard (HD) with decontaminants in organic 

matrices 

Holden, Ian; Rothery, Emma; Govan, Norman 

Dstl, Detection, Porton Down, United Kingdom 

Mustard gas (HD, bis(2-chloroethyl)sulfide) was first used in World War I 

as a potent vesicant. Although its production is now banned in countries 

which are signatories to the Chemical Weapons Convention there are 

still a large number of legacy issues associated with it. These involve 

both environmental and personal contamination. In order to develop new 

decontaminants it is necessary to monitor non-invasively the processes 

of the associated chemical reactions in a variety of media. This presents 

a number of technical problems in that both products and reactants 

have different polarities and volatilities and are present in a complex 

background matrix making monitoring by conventional 1 H and 13 C NMR 

and/or hyphenated MS techniques impractical. The INADEQUATE NMR 

technique in various forms has been primarily and extensively used 

since 1980 to determine the carbon backbone of organic structures. 

We have utilised INADEQUATE NMR spectroscopy to suppress the 

natural abundance 13 C signals from a variety of organic matrices, 

in order to monitor the fate of dilute solutions (ca. 1-5%) of labelled 

[ 13 C- 13 C]bis(2-chloroethyl)sulfide in a number of decontaminants. The 

course of the reactions of labelled mustard with chlorine dioxide in 


the complex decontaminant mixture F32, together with the personal 

decontaminant RSDL* (2,3 butanedione 2-oxime potassium salt in 

monomethylated polyethylene glycol) are described. The 1D and 2D 

INADEQUATE techniques give excellent suppression of the background 

signals permitting tentative product identification especially if used in 

conjunction with other 2D NMR methods e.g. HMQC. They provide the 

basis for a general protocol for studying the fate of mustard in any liquid 

preparation. *Reactive Self-Decontamination Lotion. 

Sm23 

Structure of an N-methylated MT-II derivative which is a highly 

potent and selective melanocortin receptor subtype 1 agonist 

Doedens, Lucas 1 ; Opperer, Florian 1 ; Beck, Johannes G. 1 ; Minying, Cai 2 ; 

Dedek, Matt 2 ; Palmer, Erin 2 ; Hruby, Victor J. 2 ; Kessler, Horst 3 

1 TU München, Department Chemie, München, Germany; 2 University of 

Arizona, Department of Chemistry, Tucson, United States; 3 Institute for 

Advanced Study at the TU München, München, Germany 

MT-II (Ac-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-NH 2 ) is a highly active 

but also unselective agonist for the melanocortin receptor subtypes 

1 and 3-5. It is well known that substitution of amide protons with 

methyl groups can result in receptor subtype selectivity and changes 

in pharmacological properties, such as metabolic stability, lipophilicity, 

enhanced potency, enhanced bioavailability and conformational rigidity. 

We chose MT-II as template for our investigations on the effects of 

N-methylation on conformation of cyclic peptides as ligands for the 

melanocortin receptors and its influence on bioavailability. Systematic 

N-methylation of the five amide bonds inside the cyclic system was 

performed yielding 2 5 = 32 differentially N-methylated analogs. 

Biological testing shows that several of these peptides still have high 

activity but they differ strongly for the individual receptor subtypes. 

The synthesized peptides were tested with respect to competitive 

binding and adenylate cyclase activation using HEK-293 cells 

expressing hMCR1, 3-5. The structure of one highly potent agonistic 

and hMCR1 selective N-methylated MT-II derivative was investigated 

by NMR spectroscopy. Based on NMR data (69 ROE derived distance 

restraints, homo- and heteronuclear coupling constants) as well as 

on restrained and free molecular dynamics a distinct and preferred 

structure could be derived for the peptide backbone. 

3 JHα-Hβ coupling constants of Asp and Lys suggest that their sidechains, 

which are connected by an amide bond, are significantly more flexible 

than the backbone. Significant line broadening of Lys H ζ further 

indicates intermediate exchange of different conformers at the end of 

the Lys sidechain. For the Nle, His and Arg sidechains, 3 J Hα-Hβ coupling 

constants indicate a high degree of flexibility with similar populations of 

the χ 1 =-60° and χ 1 =180° rotamers. The 3 J Hα-Hβ coupling constants of 

Trp indicate a strong preference of χ 1 =-60° (p=80%) which seems to 

be favoured due to hydrophobic clustering of the aromatic sidechain to 

the N-methyl groups of His and Arg. 

Sm24 

A Vanadium Compound with promising anti-diabetic properties: 

chemical and biochemical studies with a pyridinone complex 

Passadouro, Marta 1 ; Carvalho, Eugenia 1 ; Faneca, Henrique 1 ; Delgado, 

Teresa 2 ; Jones, John 1 ; Tomaz, Ana Isabel 3 ; Gonçalves, Gisela 3 ; Geraldes, 

Carlos F.G.C. 4 ; Costa Pessoa, João 3 ; Castro, M. Margarida 4 

1 University of Coimbra, Center for Neurosciences and Cell Biology, 

Coimbra, Portugal; 2 University of Coimbra, Biochem.Dept; CNC, 

Coimbra, Portugal; 3 Technical University of Lisbon, Lisbon, Portugal; 

4 University of Coimbra, Biochem.Dept.; CNC, Coimbra, Portugal 

The importance of Vanadium Compounds (VCs) has increased in the 

last years due to their pharmacological properties. In particular, their 

potential use as oral insulin mimetics 1 has been demonstrated by in 

vivo and ex vivo studies, as well as in clinical trials. In this work we 

report some studies with a VC containing a pyridinone ligand, the bis- 

[3-hydroxy-1,2-dimethyl-4-pyridonate]oxovanadium (IV), V IV O(dmpp) 2 , 

which shows promising anti-diabetic capacity. 

The structural characterization and solution behaviour of the main V(V) 

species present in different aerobic aqueous solutions, at physiological 

pH, were studied, showing some favourable properties concerning 

solubility and stability. The interaction of this VC with human transferrin 

was also investigated using 51 V NMR and EPR spectroscopy to check if 

this plasma protein may facilitate the entry of this VC in the cells. 

The uptake of this VC by human erythrocytes was studied using 51 V and 

1 H NMR and EPR spectroscopy 2 and its oxidative stress was evaluated 

by measuring its effect on the pentose phosphate pathway flux using 

[2- 13 C]-glucose and 1 H NMR spectroscopy 3 . 

Other biochemical studies were also carried out concerning glucose 

uptake by primary cultures of rat adipocytes, using a radioactive assay, 

to prove its capacity to withdraw insulin resistance in these cells. It was 

observed that in a nontoxic concentration, as confirmed by the Alamar 

Blue test, this VC increases glucose uptake relative to basal values in 

the absence of insulin and enhances insulin action. It was also observed 

that it reverts the action of immunosuppressants. Thus, this VC may 

represent a major candidate as a powerful drug to treat diabetes and 

other metabolic disorders. 

References 

1. D. C. Crans, J. J. Smee, E. Gaidamauskas, L. Yang Chem. Rev. 2004, 

104: 849-902. 

2. T. Delgado, A.I. Tomaz, I. Correia, J. Costa Pessoa, J. Jones, C. 

F.G.C.Geraldes and M.M.Castro, J. Inorg.Biochem., 2005, 99: 2328- 

2339 

3. T.Delgado, M. Castro, C. Geraldes, J. Jones., Magn.Reson.Med. 

2004, 51: 1283-1286 

Acknowledgements The authors thank the support from FEDER and FCT, 

Portugal, POCI 2010, Project PPCDT/QUI/56949/2004 


Sm25 

Three-dimensional NMR spectroscopy of organic molecules 

Misiak, Maria; Kozminski, Wiktor 

University of Warsaw, Faculty of Chemistry, Warsaw, Poland 

Multidimensional NMR Spectroscopy can be regarded as a powerful 

technique used for studies of molecular structure and dynamics of 

complex organic compounds. 

Here we present the application of 3D experiments, which are 

based on random sampling of the evolution time space followed by 

Multidimensional Fourier Transform (MFT). This approach, applied at 

first to the strychnine molecule, enables one to acquire 3D spectra in 

reasonable experimental time and allows We also show that by the 

application of 3D NMR experiments we were able to do the complete 

1 H and 13 C spectral assignment of natural abundance prenol-10, what 

was impossible earlier using 1D and 2D techniques because of the 

complexity of the spectra. 

Furthermore, we show here an application of 3D NMR experiments 

for measurement of heteronuclear coupling constants of organic 

compounds yielding complex spectra. 

The interpretation of 3D HSQC-TOCSY spectra with E.COSY-type 

multiplets allowed us to evaluate heteronuclear coupling constants of 

strychnine with high accuracy, what is difficult employing 2D methods 

owing to signal overlap and impossible using conventionally recorded 

3D NMR spectra in the reasonable time of an overnight experiment. 

[1] Kazimierczuk K, Kozminski W, Zhukov I, J. Magn. Reson. 179, 323 

(2006) 

[2] Kazimierczuk K, Zawadzka A, Kozminski W, Zhukov I. J. Biomol. 

ABSTRACTS POSTER


NMR. 36, 157 (2006) 

[3] Kazimierczuk K, Zawadzka A, Kozminski W, Zhukov I. J. Magn. 

Reson. 188, 344 (2007) 

[4] Misiak M, Kozminski W, Magn. Reson. Chem, 45, 171 (2007) 

[5] Kazimierczuk K, Zawadzka A, Kozminski W. J. Magn. Reson. 192, 

123 (2008) 

[6] Kazimierczuk K, Zawadzka A, Kozminski W, Zhukov I. J. Am. Chem. 

Soc. 130, (16) 5404 (2008) 

[7] Misiak M, Kozminski W, Magn. Reson. Chem. 47, 205 (2009) 

Sm26 

Ecdysteroids: Compounds with promising biological activity - 

structural determination 

Takács, Mária 1 ; Simon, András 2 ; Tóth, Gábor 2 ; Liktor-Busa, Erika 3 ; Tóth, 

Noémi 3 ; Báthori, Mária 3 ; Gergely, András 1 

1 Semmelweis University, Department of Pharmaceutical Chemistry, 

Budapest, Hungary; 2 Budapest University of Technology and Economics, 

Department of Inorganic and Analytical Chemistry, Budapest, Hungary; 

3 University of Szeged, Department of Pharmacognosy, Szeged, Hungary 

Ecdysteroids are know to be moulting and metamorphosis hormones 

of insects. Compounds structurally related to anthropods’ steroid 

hormones occur in wide range of, and multiple structural variations 

in plants. Pharmacological studies have revealed that ecdysteroids 

influence many physiological functions in a positive way, not being 

toxic to mammals. Their most pronounced effect on mammals 

is a stimulation of protein synthesis without adverse androgenic, 

antigonadrotopic, or thymolytic side-effects. Recently, biotechnology 

started to employ in ecdysteroids as powerful inducers for gene-swich 

systems, that can be promising regarding controlled systems for human 

therapies. 

Compounds to be presented are the following: three new natural 

ecdysteroids, 22-dehydro-20-deoxy-ajugasterone C (1), 1-hydroxy-22deoxy-20,21-didehydro-ecdysone 

(2) and 22-deoxy-20,21-didehydroecdysone 

(3) were isolated from the methanol extract of the roots of 

Serratula wolffii. Two new, 2-deoxypolypodine B 25β-D-glucoside (4), 

2-deoxypolypodine B 22-glucoside (5) and three known, 2-deoxy- 

20-hydroxyecdysone 3-glucoside (6), 2-deoxypolypodine B 3β-Dglucoside 

(7), integristerone A 20,22 monoacetonide (8), ecdysteroids 

were identified from the herb of Silene viridiflora. The structures 

of compounds 1-8 were established by extensive spectroscopic 

techniques, including one- and two-dimensional NMR methods and 

mass spectrometry. 

Due to our examinations, Serratula wolffii is proven to be a valuable 

source of the 11α-OH hydroxy ecdysteroids, that group is important 

for the manifestation of anabolic activity of ecdysteroids. Plants 

biosynthetise several ecdysteroid-glucosides, outstandingly Silene 

species. As ecdysteroid-glucosides are of good water-solubility, the 

study of their pharmacological effects is especially available. 

Sm27 

Investigation of solution state structure of thiosemicarbazone 

derivatives by NMR and DFT methods 

Piculjan, Katarina 1 ; Hrenar, Tomica 1 ; Smrecki, Vilko 2 ; Rubcic, Mirta 1 ; 

Cindric, Marina 1 ; Novak, Predrag 1 

1 Faculty of Science, University of Zagreb, Chemistry, Zagreb, Croatia; 

2 Rudjer Boskovic Institute, Chemistry, Zagreb, Croatia 

Salicylaldehyde thiosemicarbazones and their metal complexes belong 

to an important class of biologically active compounds (anticancer, 

antivirial, antibacterial, antiinflammatory and antifungal activity) [1]. They 

can exist in several tautomeric forms e.g. hydroxy-thione, hydroxy-thiol, 

keto-thione and keto-thiol with both intra- and inter-molecular hydrogen 

98 

bonds. 

X-ray structural analysis has confirmed the existence of intra-molecular 

hydrogen bonds in different salicylaldehyde thiosemicarbazone 

derivatives [2,3]. Bioactivity is closely related to molecular structure 

which is governed by the presence of hydrogen bonds. Therefore, 

the aim of our study was to investigate the influence of solvent and 

substituents on molecular conformation and structure of hydrogen 

bonds in salicylaldehyde thiosemicarbazones by combining NMR and 

DFT methods [3-5]. Solvents of different polarities, i. e. of different 

proton donor and acceptor abilities were used (chloroform, acetone, 

methanol, dimethyl sulfoxide). Also, the effect of substituting OH with 

OMe group in salycilaldehyde residue, on the overall thiosemicarbazone 

structure and tautomerism was explored. Two-dimensional NOESY 

spectra indicated conformational changes in solution with respect to the 

structure observed in solid state owing to a relatively low barrier of the 

rotation arround N-N single bond thus enabling a molecule to posses a 

higher conformational flexibility in solution. The results presented here 

can help in a better understanding of the role hydrogen bonds can play 

in bioactivity of related thiosemicarbazone derivatives and their metal 

complexes. 

Sm28 

Covariance spectroscopy and old-fashioned NMR experiments – 

A comparative study 

Aspers, Ruud; Jaeger, Martin 

Schering-Plough Corporation/Schering-Plough Research Institute, 

Department of Medicinal Chemistry, Oss, Netherlands 

Structure elucidation and structure verification of small molecules 

are major tasks in pharmaceutical analysis. As being part of today’s 

standard procedures, such analyses are often subject to severe time 

constraints. In addition, limited amount of sample is dedicated to 

analyses, causing an increase in experimental and cycle time. While 

the latter can be significantly reduced by using appropriate correlation 

techniques, the experiment time expands along with experimental 

complexity and decreased sensitivity. 

NMR experiments that rely on complex but straight-forward pulse 

sequences and coherence pathways are well established and form the 

toolbox of every NMR laboratory[1]. In recent years, alternative routes 

have been discovered [2], explored [3,4] and made available [5]. One 

of these methods was baptized Covariance Spectroscopy and is based 

on a mathematical transformation of two different types of spectra into 

one, containing the information of both. On top of it, the transformation 

correlates the formerly separate information. 

In the current study, steroids as examples of increasingly crowded 

spectra are investigated by means of covariance spectroscopy and 

known, but time consuming and material requiring NMR experiments 

such as HSQC-TOCSY and 13C,13C-INADEQUATE. The information 

obtained from both methods are compared. The similarities and 

differences between the two apporoaches will be demonstrated. 

The pros and cons of the experiments will be discussed in terms of 

utilization under standard conditions. 

1. S. Berger 250 and more NMR experiments, Wiley-VCH, Weinheim, 

200x. 

2. F. Zhang and R. Bruschweiler, J. Am. Chem. Soc., 126, 13180 

(2004). 

3. K. A. Blinov, N. I. Larin, M. P. Kvasha, A. Moser, A. J. Williams, and G. 

E. Martin, Magn. Reson. Chem., 43, 999 (2005). 

4. K. A. Blinov, A. J. Williams, B. D. Hilton, P. A. Irish, and G. E. Martin, 

Magn. Reson. Chem., 45, 544 (2007). 

5. http://www.acdlabs.com/publish/publ07/enc2007_indirect_ 

covariance_nmr.html 


Solid State NMR 

Sn10 

Supramolecular association leading to hydro/organogelation: 

A solid state NMR study 

Nonappa, ; Lahtinen, Manu; Behera, Babita; Kolehmaninen, Erkki; 

Kauppinen, Reijo 

University of Jyväskylä, Department of Chemistry, Jyväskylä, Finland 

Because of their unique ability to immobilize and encapsulate a variety 

of solvents, low molecular mass organo/hydrogelators (LMOGs) has 

gained enormous attention in recent years. 1 Structural diversity of 

spontaneously self-assembled physical gels in the nanometre regime 

make them excellent media for potential applications in the fields of 

drug delivery, tissue engineering, immobilization of protein/peptide 

arrays, medicine, biomimetics and nanoscience. 2 Unusual gelation 

of organic fluids by simple esters of bile acid was recently reported. 3 

A number of experimental evidences suggest a close resemblance 

between the xerogels and bulk solid. However, several attempts towards 

the crystallization of gelators have been unsuccessful. Therefore, a 

clear understanding about the non-covalent interactions leading to 

the gelation remained partly undisclosed. This prompted us to use 13 C 

CP-MAS NMR as a tool to shed light into the nature and molecular 

packing of bulk solid, xerogels, and organo/hydrogels. Interestingly, 

13 C CP-MAS of the gelators displayed unique spectral patterns, while 

non-gelators either showed more simple spectral patterns or resulted 

in the formation of inclusion complexes, solvates and polymorphs. The 

experimental results obtained in these studies lead us to answer a 

number of questions in the field of physical gels (one of the burgeoning 

branch of supramolecular chemistry) which otherwise remained 

unanswered. The results obtained from these experiments will be 

discussed in the presentation. 

References: 

1. (a) N. M. Sangeetha, U. Maitra, Chem. Soc. Rev. 2005, 34, 821; (b) 

Molecular Gels: Materials with Self-Assembled Fibrillar Networks, ed. R. 

G. Weiss, P. Terech, Springer, Netherlands, 2006. 

2. (a) S. Murdana, Expert Opin. Drug Delivery, 2005, 2, 489; (b) A. 

Valkonen, M. Lahtinen,; E. Virtanen,; S. Kaikkonen,; E. Kolehmainen, 

Biosens. Bioelectron. 2004, 20, 1233; 

3. Nonappa.; U. Maitra. Soft Matter, 2007, 3, 1428. 

Sn11 

Low-power solid-state NMR experiments for resonance 

assignments under fast magic-angle spinning 

Vijayan, Vinesh 1 ; Demers, Jean-Philippe 1 ; Becker, Stefan 1 ; Biernat, 

Jacek 2 ; Mandelkow, Eckhard 2 ; Lange, Adam 1 

1 Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany; 

2 Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany 

Micro-crystalline globular proteins, amyloid fibrils, and membrane 

proteins can now be routinely studied using solid-state NMR techniques. 

This was made possible in part due to the development of 2D and 3D 

homonuclear and heteronuclear experiments that correlate 13 C and 

15 N spins for resonance assignment and also for obtaining long-range 

distance restraints in structure elucidation. 

Remarkable development in magic angle spinning (MAS) probe 

technology also contributed to this success. Now, a new generation of 

commercially available 1.3 mm probes can reach above 60 kHz of MAS. 

This allows for more efficient averaging of strong dipolar couplings 

hence providing better resolution in highly crowded protein spectra. 

On the other hand, fast spinning also reduces the effectiveness of 

many of the routinely used NMR experiments for obtaining resonance 

assignments. 

Here, we present a complete set of solid-state NMR experiments 

sufficient for protein resonance assignments under fast MAS (> 60 kHz) 

including 15 N- 13 C correlation experiments. The low-rf field requirements 

of our experiments make the approach well suited for the investigation 

of temperature-sensitive biomolecules without the risk of sample 

heating and degradation. 

We present NCA, N(CO)CA, and 13 C- 13 C correlation spectra at 60 

kHz MAS that were recorded on less than 1 mg of [ 13 C, 15 N] isotope 

labeled sample. We also demonstrate that our approach can be readily 

performed on protein samples in which the 1 H T 1 relaxation times are 

shortened by means of paramagnetic metal ions. Here, the reduced 

recycle delay requires the use of NMR sequences with a reduced power 

deposition as described in this work. 

Shown applications include micro-crystalline ubiquitin and Alzheimer’slike 

paired helical filaments from the core domain of tau protein. 


Sn12 

Time-resolved solid state NMR in studies of biocatalytic 

transformations: application and perspectives 

Cherepanov, Alexey 

Johann Wolfgang Goethe-University, BMRZ - Center for Biomolecular 

Magnetic Resonance, Frankfurt am Main, Germany 

The presentation will discuss the application of time-resolved solid 

state NMR spectroscopy in studies of molecular kinetics of enzymatic 

reactions. Two methods of starting the reaction will be reviewed, 

phototriggering and direct mixing of the reactants followed by a rapid 

freezing. Preparation of the samples and instrumentation details will be 

described. The presentation will demonstrate the NMR ability to detect 

changes of the environment around the observed nuclei as the chemical 

process advances over the energy barrier along the reaction coordinate. 

Presented results at cryogenic temperatures will demonstrate the 

capacity of NMR spectroscopy to monitor the processes of makingand-breaking 

of chemical bonds in real time. Within the framework of 

structural NMR studies, attention will be paid to the molecular details of 

several biochemical reactions: Mg 2+ -assisted adenylyl transfer reaction 

catalyzed by T4 DNA ligase, intramolecular redox reaction of caged ATP 

and binding of azide to metmyoglobin from horse heart. 

Sn13 

Proton double-quantum of solid polymers in inhomogeneous 

fields 

Adams, Alina 

ITMC, RWTH Aachen University, Macromolecular Chemistry, Aachen, 

Germany 

Recently, the possibility of exciting and detecting proton doublequantum 

NMR coherences of hard polymers in inhomogeneous fields 

was shown for the first time [1]. For this, pulse sequences which 

partially refocus the effect of the field inhomogeneities on the spin 

system evolution and generate double-quantum build-up curves were 

used. The theoretical justification of the method was based on a simple 

two spin ½ system. The performance of the same pulse sequence was 

tested also with a high field solid-state NMR spectrometer working at 

500 MHz. The methodology was validated by measurements performed 

for a series of covalently cross-linked polymer networks based on 

random copolymers of L,L-dilactide and diglycolide with potential 

medical applications. These systems exhibit strong dipolar interactions, 

and due to their insolubility, solid state NMR techniques are required. 

The method was applied to investigate the effect of the chain length on 

the chain dynamics via the values of the residual dipolar couplings and 

ABSTRACTS POSTER


to follow for the first time the kinetics of UV curing for the same type 

of polymer network. Good agreement with the values yield by high field 

NMR was obtained. However, the main drawback of the implemented 

method is related to the poor signal to noise ratio and therefore a very 

long measuring time is needed for quantitative results. 

Further on, in this contribution, it will be shown in a first step how 

the above methodology can be further on extended for dramatically 

improvement of the signal to noise ratio by using multi-echo acquisition. 

This new approach makes the method attractive for the characterization 

of various kind of processes in real time in a truly non-destructive way. 

Moreover, it will be demonstrated that the same approach can be used 

to estimate domain sizes in heterogeneous polymers. At the end, the 

advantages and disadvantages of the method as well as the potentially 

new applications opened up by the presented results will be discussed. 

[1] A. Adams, Proton double-quantum of hard polymers in 

inhomogeneous fields (to be submitted). 

Sn14 

Aggregation behaviour of rod-coil copolymers based on 

oligoaramides - a solid state NMR study 

Bohle, Anne; Brunklaus, Gunther; Spiess, Hans-Wolfgang 

Max Planck Institute for polymer research, Mainz, Germany 

Rod-coil copolymers constitute a rather interesting class of diblock 

copolymers due to their favorable aggregation behavior. Particularly, 

compared to coil-coil copolymers, rod-coil copolymers aggregate on 

the nanometer scale, thus offering potential applications for the tailored 

design of supramolceular architectures. 

Since such architectures rarely form crystallites suitable for singlecrystal 

X-ray analysis, we employed contemporary high-resolution 

solid-state NMR at very fast magic-angle spinning to investigate 

structure and dynamics of polyethyleneglycol-oligo-p-benzamide blockcopolymers. 

Insights into both the local packing (e.g. pi-stacking) and 

nature of the hydrogen-bonding network were derived from 2D- 1 H- 1 

H and 1 H- 13 C multinuclear correlation experiments, while 13 C- and 

15 N CP-MAS NMR measurements revealed structural changes of the 

oligo-p-benzamide rods upon block-copolymer formation. The phase 

transition behavior and dynamics of the block-copolymer was studied 

via 1 H- 13 C RE-REDOR and 1 H- 13 C REPT-HDOR measurements at 

variable temperatures and discussed with respect to the local structure. 

[1] R. Abbel, H. Frey, D. Schollmeyer, A. F. M. Kilbinger, Chem. Eur. J. 

2005, 11, 2170-2176. 

[2] T. W. Schleuss, R. Abbel, M. Gross, D. Schollmeyer, H. Frey, M. 

Maskos, R. Berger, A. F. M. Kilbinger, Angew. Chem. Int. Ed. 2006, 45, 

2969–2975. 

Sn15 

Structural insights into the Ure2p full-length prion protein by 

solid-state NMR spectroscopy 

Loquet, Antoine 1 ; Bousset, Luc 2 ; Gardiennet, Carole 1 ; Sourigues, 

Yannick 2 ; Habenstein, Birgit 1 ; Schuetz, Anne 3 ; Wasmer, Christian 3 ; Meier, 

Beat 3 ; Melki, Ronald 2 ; Böckmann, Anja 1 

1 IBCP-CNRS, Lyon, France; 2 LEBS-CNRS, Gif-sur-Yvette, France; 3 ETH 

Zurich, Zurich, Switzerland 

Protein misfolding and subsequent aggregation is the hallmark of 

over 30 diseases termed “conformational” diseases. Although actively 

studied, the fundamental mechanism of the misfolding of a variety of 

polypeptides leading to their aggregation and associated diseases is 

far from being understood. A subset of neurodegenerative illnesses 

is linked to the aggregation of an infectious protein termed prion. 

100 

Prions assemble in vitro into fibrillar structures that are thought to be 

infectious through their ability to grow by incorporating the constitutive 

form of the prion molecules. The heterogeneity of prion high molecular 

weight oligomers and their molecular mass make them not suitable for 

structural studies at atomic resolution by classical methods as x-ray 

crystallography and solution NMR. 

The yeast prion Ure2p is a two-domain protein. The compactly folded 

C-terminal domain spans residues 94-354 and the crystal structure 

of this C-terminal domain was solved by x-ray crystallography. The 

N-terminal, critical for prion propagation ranging from amino acid 1 to 

93, is rich in asparagine and glutamine residues and insoluble in its 

native form. In a manner similar to other prions, full-length Ure2p fibrils 

assembled under physiological conditions share several morphological 

and tinctorial features with amyloids. However, the fibrils lack the 

canonic X-ray diffraction pattern and the specific absorbance in infrared 

light that characterizes amyloids. 

We here present solid-state NMR experiments on full-length Ure2p 

yeast prion fibrils which demonstrate that fibrils formed under nearphysiological 

conditions have a mostly well-ordered and well-defined 

atomic structure leading to highly resolved NMR spectra. Data 

presented include measurements on the prion and globular domains 

in isolation, as well as in their natural context in native-like full-length 

Ure2p fibrils. We show that the structure of the C-terminal domain 

remains intact and displays a near-crystalline order within the fibrils, 

compatible with a model where the globular domain forms an integral 

part of the fibril. For the prion domain, the spectra reflect disorder 

suggesting structural inhomogeneity. 

Sn16 

Structure of full-length αB-crystallin determined with MAS NMR 

van Rossum, Barth-Jan 1 ; Jehle, Stefan 1 ; Markovic, Stefan 1 ; Diehl, 

Anne 1 ; Rehbein, Kristina 1 ; Rajagopal, Ponni 2 ; Klevit, Rachel 2 ; Oschkinat, 

Hartmut 1 

1 Leibniz-institute of Molecular Pharmacology (FMP), Structural Biology 

Programme, Berlin, Germany; 2 University of Washington, Department of 

Biochemistry, Seattle, United States 

Magic-angle spinning solid state NMR spectroscopy is potentially able 

to deliver high resolution information on large biomolecular complexes 

like membrane proteins, dynamical oliomers, proteins associated with 

the cytoskeleton, and protofibrils. We have studied the structure of 

full-length αB-crystallin. αB-crystallin belongs to the family of small 

heat-shock proteins and is the major protein constituent of the human 

eye-lense. It builds large polydisperse, dynamic oligomers. The building 

block of the oligomers is a dimer formed by two conserved crystallin 

domains; the oligomerization is driven by interactions between the 

N- and C-termini and neighbouring dimers. Due to its polydisperse 

character and due to the size of the oligomers, αB-crystallin has 

escaped high-resolution structural studies for many years. Using 

MAS NMR, we now have been able to determine the structure of the 

crystallin domain in full-length αB-crystallin oligomers. Moreover, from 

special preparations using complementary isotope labelling, we could 

map out the dimer interface and calculate a first dimer structure. 

Detection of contacts between the termini and dimer provided first 

insights into the structure of the oligomer. We also report on studies on 

a mutant, R120G, which is known to induce cataract. 


Sn17 

Phosphonic acid based ionomers as fuel cell membranes 

Fassbender, Birgit; Jimenez, Lucia; Klapper, Markus; Brunklaus, 

Gunther; Spiess, Hans-Wolfgang 


Proton conducting fuel cell membranes based on perfluorosulfonic 

acid polymers (i.e. NAFION ® ) are abundantly used in industry [1]. 

Due to limitation of the operating temperature below the dew point of 

water, these polyelectrolyte exchange membranes (PEM) face several 

problems, such as efficiency-limiting water and heat management 

and poor carbon monoxide tolerance [2]. Increasing the operating 

temperature above 100°C may overcome these drawbacks, but proton 

transport is rather difficult in a “dry” membrane [3]. Nevertheless, 

high proton conductivities (T>100°C) are obtained upon addition 

of amphoteric liquid molecules. In that case, leaching of the mobile 

molecules, e.g. due to water produced when operating fuel cells, 

causes severe problems [2]. 

In this work, proton conducting features and proton dynamics of 

ionomers with covalently bound phosphonic acids were studied using 

solid state 1 H, 2 H and 31 P MAS NMR, respectively. Therefore, different 

geometrical structures with varied weight fractions of phosphonic acid 

groups and either as aromatic or perfluorinated scaffold structure were 

obtained via different synthetic routes. 

In particular, (apparent) activation energies of proton mobility obtained 

from temperature-dependent 1 H NMR measurements of the different 

ionomers are compared with macroscopic results derived from 

impedance spectroscopy. In addition, static and MAS 2 H NMR spectra 

were recorded to reveal the kind of motion involving the hydrogenbonded 

protons of the phosphonic acid group. 

Furthermore, combining TGA results with 31 P VT NMR measurements 

allowed for insights into both the mechanism and amount of selfcondensation 

of phosphonic acid groups. 

[1] de Bruijn, F.A., Makkus, R.C., Mallant, R. K., Janssen, G.J., Adv. in 

Fuel Cells, 2007, 1, 235-336 

[2] Steininger, H., Schuster, M. Kreuer, K.D.; Kaltbeitzel, A., Bingoel, B., 

Meyer, W. H.; Schauff, S., Brunklaus, G., Maier, J., Spiess, H. W., Phys. 

Chem. Chem. Phys. 2007, 9, 1764-1773 

[3] Kaltbeitzel, A., Schauff, S., Steiniger, H., Bingoel, B., Brunklaus, G., 

Meyer, W., Spiess, H.W., Solid State Ionics 2007, 178, 469-474 

Sn18 

Solid-state NMR studies of lipid anchored oligonucleotides as 

nanotechnological building blocks 

Bunge, Andreas 1 ; Liebscher, Jürgen 2 ; Herrmann, Andreas 2 ; Huster, 

Daniel 1 

1 University of Leipzig, Leipzig, Germany; 2 Humboldt University, Berlin, 

Germany 

For the development of surface functionalized bilayers we have 

synthesized lipophilic oligonucleotides to combine the molecular 

recognition mechanism of nucleic acids and the self-assembly 

characteristics of lipids in planar membranes. We tested several 

nucleoside monomers with regard to their membrane binding 

characteristics and their suitability as builiding blocks for oligonucleotide 

synthesis using 2 H and 31 P NMR. Using these building blocks, lipophilic 

oligonucleotides consisting of 20 to 25 nucleotides and two lipophilic 

anchors such as α-tocopherol moieties or simple hexadecyl chains as 

lipophilic anchor were synthesized using solid-phase methods with 

phosphoramadite strategy. The interaction of the water-soluble lipophilic 

oligonucleotide with vesicular lipid membranes and its capability to 

bind complementary DNA strands was studied using complementary 

methods such as NMR, EPR, DSC, fluorescence spectroscopy and 

fluorescence microscopy. These oligonucleotides inserted stably into 

preformed membranes from the aqueous phase. Thereby, no significant 

perturbation of the lipid bilayer and its stability was observed. However, 

the non-lipidated end of the oligonucleotide is exposed to the aqueous 

environment, relatively mobile, and free to interact with complementary 

DNA strands. Binding of the complementary single-stranded DNA 

molecules is fast and accomplished by formation of Watson-Crick base 

pairs, which was confirmed by 1 H NMR chemical shift analysis and 

fluorescence resonance energy transfer. The molecular structure of the 

membrane bound DNA double helix is very similar to the free double 

stranded DNA. Further, the membrane bound DNA double strands also 

undergo regular melting. Finally, in raft-like membrane mixtures, the 

lipophilic oligonucleotide was shown to preferentially sequester into 

liquid-disordered membrane domains. 

References: 

A. Kurz et al. Angew. Chem. Int. Ed. 45 (2006) 4440-4444 

A. Bunge et al. Langmuir 23 (2007) 4455-4464 

N. Brodersen et al. Eur. J. Org. Chem. 36 (2007) 6060-6069 

O. Kaczmarek et al. Eur. J. Org. Chem. 37 (2008) 1917-1928 


Sn19 

Structure determination of N-acetyl-glucosamine with 15 N/ 13 C 

multiple isotope labeling by SPECIFIC REDOR spectroscopy 

Lu, Yi-Zhi 1 ; Chia, Winlong 1 ; Tzou, Der-Lii 2 

1 Fu-Jen Catholic University, Department of Chemistry, Taipei, Taiwan; 

2 Institute of Chemistry, Academia Sinica, Nankang Taipei, Taiwan 

We introduce a modified REDOR spectroscopy, termed as SPECIFIC 

REDOR, that is suitable for determining internuclear distances in 

multiple 15N/ 13C isotope labeled compounds. In this technique, a 

double CP scheme, comprising 1H/ 15N and 15N/ 13CPs, is combined with 

REDOR for specific 15N/ 13C dipolar interaction observation. Given that 

15 13 N/ C coherence transfers are extremely sensitive to the experimental 

parameters, specific distance measurements in 15N/ 13C multiple isotope 

labeled N-acetyl-glucosamine (GlcNAc) are achievable by appropriate 

setting of the rf field strengths or the MAS frequency. We report here 

that the internuclear distances in GlcNAc determined by SPECIFIC 

REDOR spectroscopy [ 15N-13CO (1.35±0.03 Å), 15N-13 α C (2.57±0.23 

1 

Å, 15N-13 β 15 13 C (2.57±0.16 Å, N- CH3 (2.64±0.15 Å] are consistent with 

1 

those determined by X-ray diffaction crystallography. Given the distance 

constraints, the molecular structure of GlcNAc has been determined 

after refinement by computer modelling simulation. 

Sn20 

Characterization of chemical and geometrical disorder in 

oxide materials using multiple quantum J-mediated solid state 

experiments 

Massiot, Dominique; Fayon, Franck; Deschamps, Michael; Hiet, Julien; 

Pellerin, Nadia; Montouillout, Valérie; Cadars, Sylvian; Véron, Emmanuel; 

Florian, Pierre 

CEMHTI CNRS UPR 3079 Université d’Orléans, Orléans, France 

Although oxide glasses appear as homogeneous and isotropic materials 

at the macroscopic or microscopic scales they are inhomogeneous 

and anisotropic at the atomic to sub-nanometer scales which remain 

challenging to characterize. The understanding of glasses properties 

or the glass forming process through the glass transition and more 

generally of their thermodynamic properties specifically depends upon 

their local structure. 

We describe a series of solid state NMR experiments that make use 

of small but detectable homo- or hetero-nuclear indirect J coupling 

to identify extended structural motifs through correlation experiments, 

spin counting experiments or multiple quantum (up to 4 or 5 quanta) 

ABSTRACTS POSTER


filtered experiments to identify Qm(nAl) silicon or aluminum sites. 

These experiments enable evidencing separately chemical disorder and 

geometric disorder in silicate or aluminosilicate glasses. 

[1] D.Massiot, F.Fayon, M.Deschamps, S.Cadars, P.Florian, 

V.Montouillout, N.Pellerin, J.Hiet, A.Rakhmatullin, C.Bessada, “Detection 

and use of small J couplings in solid state NMR experiments” Submited 

[2] J.Hiet, M.Deschamps, N.Pellerin, F.Fayon, D.Massiot “Probing 

chemical disorder in glasses using silicon-29 NMR spectral editing” 

Submited 

[3] S.K.Lee, M.Deschamps, J.Hiet, D.Massiot, S.Y.Park “Connectivity 

and proximity between quadrupolar nuclides in oxide glasses: Insights 

from through-bond and through-space correlations in solid-state NMR” 

J. Phys. Chem. B asap 2009 

[4]P.Florian, F.Fayon, D.Massiot “2J Si-O-Si scalar spin-spin coupling in 

the solid-state: the case of crystalline and glassy wollastonite CaSiO3” 

J. Phys. Chem. C 113 2562-2572 2009 

[5]D.Massiot, F.Fayon, V.Montouillout, N.Pellerin, J.Hiet, C.Roiland, 

P.Florian, J.P.Coutures, L.Cormier, D.R.Neuville, “Structure and dynamics 

of Oxyde Melts and Glasses: a view from multinuclear and high 

temperature NMR” J. Non-Cryst. Solids 354 249-254 2008 

Sn21 

Single crystal micro MAS NMR: Quadrupole tensor orientation 

and structural disorder 

van Eck, Ernst RH; Vasa, Suresh K; Janssen, Hans WG; Kentgens, Arno 

PM 

Radboud University Nijmegen, Solid State NMR - IMM, Nijmegen, 


Currently most structural investigations using solid-state NMR are 

performed on powdered samples, spinning at the magic-angle. As has 

already been shown in the early days of NMR studying single-crystals 

can supply additional information as it gives the orientation of the tensor 

interactions and therefore the orientation of the molecular moieties in 

the crystal. Generally, however, single crystals of the sufficient size to do 

NMR measurements are not available. 

Here we show how our recently developed µ-MAS (Magic Angle 

Spinning) technology can be successfully employed for determining 

quadrupolar tensors and their orientation in a small crystal. Having 

established a method to obtain the quadrupolar tensor orientation in 

a larger size crystal (1.1 x 1.1 x 6 mm) of NaNO 3 , using a standard 

2.5 mm MAS probe, we demonstrate the feasibility of this approach 

for the micro MAS set up. A 150 x 150 x 800 µm crystal of sodium 

nitrate was spun at 12.5 kHz and the Euler angles of the quadrupole 

tensor are obtained. Furthermore, for samples with inherent disorder, 

the study of single-crystals allows one to directly access this disorder. 

This is illustrated with the zeolite mesolite where multiple quantum MAS 

(MQMAS) data obtained from a powdered sample are compared with 

that of a micro sized single crystal. 

102 

Sn22 

17 O Solid-state NMR and first-principles calculations of 

amorphous sodium phosphates 

Vasconcelos, Filipe 1 ; Cristol, Sylvain 1 ; Paul, Jean-François 1 ; Charpentier, 

Thibault 2 ; Amoureux, Jean-Paul 1 ; Montagne, Lionel 1 ; Mauri, Francesco 3 ; 

Delevoye, Laurent 1 

1 Unité de Catalyse et de Chimie du Solide, Villeneuve d’Ascq, France; 

2 CEA, IRAMIS, Service de Chimie Molculaire, Gif-sur-Yvette, France; 

3 Institut de Minéralogie et Physique des Milieux Condensés, Paris, 

France 

If 31 P is largely used for structural characterisation of crystalline and 

amorphous phosphates through Solid- State MAS NMR, very few 

17 O MAS NMR studies are reported in the literature due to intrinsic 

experimental difficulties, some of them being related to the quadrupolar 

properties of this nucleus. Our strategy is to associate crystallographic 

data, MAS NMR results and first-principles calculation in order to 

better understand the influence of 17 O local environment onto the NMR 

parameters (chemical shift, quadrupolar parameters). The calculations 

were performed using the GIPAW algorithm, developed by Pickard and 

Mauri [1], which gives access to the NMR chemical shielding for all 

nuclei of a given structure. This method improves previous strategies 

based on first-principles calculation by considering the symmetrical 

translation properties in crystals [2]. 

In this presentation, we will report on the assignment of high-field 

(18.8 T) 17 O MAS and 3QMAS spectra by the use of first-principles 

calculations for sodium metaphosphate (NaPO 3 ) and nitrate phosphate 

(“NAPON”) glass. Theoretical glass structures were obtained by both 

classical molecular dynamics (force-field) and quantum molecular 

dynamics (Born-Oppenheimer MD). The calculated parameters, 

quadrupolar constant (C Q ), quadrupolar asymmetry (ηQ) and the 

isotropic chemical shift (δiso) will be discussed and compared to those 

deduced experimentally. Original description of obtained distributions 

will also be presented following a “Czjzek approach”. 

[1] C. J. Pickard and F. Mauri , Phys. Rev. B (2001) 63, 245101 

[2] F. Vasconcelos et al., Inorg. Chem. (2008) 47, 7327 

Sn23 

Determination of Si-Al connectivities in zeolites with 2D Al→Si 

RAPT CPMAS CPMG HETCOR NMR 

Kennedy, Gordon 1 ; Wiench, Jurek 2 ; Pruski, Marek 2 

1 ExxonMobil Research and Engineering, Annandale, NJ, United States; 

2 Ames Laboratory, Iowa State University, Ames, IA, United States 

Solid state NMR spectroscopy is recognized as an integral tool in the 

structural characterization of zeolites. It provides a unique view of 

local T-site environments, including the microdistribution of framework 

atoms, reactive sites, and connectivities. The development and 

application of multidimensional solid state NMR techniques, such as, 

INADEQUATE, MQMAS, and MQ-selected heteronuclear correlation 

(HETCOR) experiments have lead to an increasingly important role in 

structure solution of zeolitic materials. These techniques have had 

many practical applications in the detailed structural characterization 

of completely siliceous zeolites and aluminophosphates. However, 

HETCOR NMR is not readily applicable to aluminosilicates due to 

reduced sensitivity. We demonstrate that the sensitivity enhancement 

obtained by the combined application of RAPT (rotor assisted population 

transfer) and CPMG (Carr-Purcell-Mieboom-Gill) is sufficient to 

efficiently measure CP-based 2D 27Al-29Si HETCOR data of complex 

microporous aluminosilicates. Using zeolite ZSM-4 as an example, we 

report high quality 2D 27Al-29Si correlation spectral data with much 

higher resolution and sensitivity (4X increase in S/N which is equivalent 

to a ~16X savings in time) than previously available. Analysis of the 2D 


data set allows complete assignment of the deceptively simple 1D 29Si 

MAS NMR spectrum into eight overlapping resonances and to map the 

Al-Si connectivities of the two inequivalent T sites. This example shows 

the potential of this technique to directly resolve Al siting and spectral 

assignment questions in this important class of materials, particularly 

those with multiple T sites. In the particular case of ZSM-4 the 

combined sensitivity and resolution enhancements allow determination 

of the existing ambiguities in spectral assignments in this material. 

Sn24 

Mg-25 ultra-high field solid state NMR and first principles 

calculations in magnesium salts 

Moudrakovski, Igor 1 ; Pallister, Peter 2 ; Ripmeester, John 1 

1 Steacie Institute for Molecular Sciences, National Research Council, 

Ottawa, Canada; 2 Carleton University, Department of Chemistry, Ottawa, 

Canada 

Magnesium has a prominent place both in geology and biology. It is the 

eighth most abundant element in the universe and the seventh most 

abundant element in the earth’s crust. However, due to the sensitivity 

problems 25 Mg remains a largely under-explored nucleus in solid state 

NMR. In this work we: 

a.) Study at ultrahigh magnetic field of 21.1T 25 Mg NMR for a number 

of previously not reported magnesium salts of known crystal structures. 

b.) Revisit and clarify the spectra of some previously reported Mgcontaining 

materials that were obtained at lower field and were either 

not sufficiently resolved, or misinterpreted. 

c.) Carry out first principles plane wave periodic system calculations 

of the 25 Mg NMR parameters (CASTEP) and compare the results to 

experimental data. The calculations produce the 25 Mg absolute shielding 

scale and give us insight into relationship between the NMR and 

structural parameters. 

At 21.1 T the effects of quadrupole interactions are reduced 

significantly and the sensitivity and accuracy in determining chemicals 

shift and quadrupole coupling parameters improve dramatically. We 

demonstrate that the chemical shift range of magnesium in diamagnetic 

compounds may approach 200 ppm. Most commonly, however, the 

observed shifts are between -15 and +25 ppm. The quadrupolar 

effects dominate the 25 Mg spectra of magnesium cations in noncubic 

environments. The chemical shift anisotropy appears to be 

rather small and only in a few cases could a contribution of the CSA 

be detected reliably. A very good correspondence has been obtained 

between the calculated shielding constants and experimental chemical 

shifts, demonstrating a good potential of computational methods in 

spectroscopic assignments of solid state NMR. 

Sn25 

Heteronuclear decoupling during symmetry-based rotorsynchronized 

sequences in magic-angle spinning solid-state 

NMR 

Scholz, Ingo; Léger, Patrick; Köneke, Stephanie; van Beek, Jacco D.; 

Meier, Beat H.; Ernst, Matthias 

ETH Zürich, Physical Chemistry, Zürich, Switzerland 

Interference effects between spin and space rotations are very 

common in solid-state NMR experiments and can be used to generate 

effective Hamiltonians that contain only selected parts of the full 

system Hamiltonian. An important class of such experiments are the 

ν ν symmetry-based CN and RNn sequences [1]. Such sequences can be 

n 

used without heteronuclear decoupling [2] at fast MAS and high rf-field 

amplitudes but very often heteronuclear spin decoupling is required. So 

far, only continuous-wave (CW) decoupling has been used. 

The problem of using multiple-pulse decoupling sequences is the 

larger number of resonance conditions due to the interference of 

the decoupling and the recoupling sequence or the sample rotation. 

We have for example experimentally and theoretically investigated 

TOBSY sequences [3] under MAS with cw and phase-alternating 

(XiX) irradiation of the protons. Multi-mode Floquet theory [4] needs 

to be applied in order to characterize all resonant and non-resonant 

phenomena. We show that in an intermediate range of decoupling 

rf-field amplitudes, XiX irradiation at resonance conditions ν r /ν XiX has 

advantages over cw irradiation in sequences. The transfer efficiency 

increases monotonically with the decoupling field strength under 

XiX decoupling while we observe strong oscillations of the transfer 

efficiency as a function of the cw decoupling amplitude. The use of XiX 

decoupling simplifies and optimizes the setup of such sequences since 

one can always use the highest possible decoupling field strength. 

References: 

1. M. H. Levitt, Encyclopedia of NMR, Vol. 9, John Wiley & Sons, Ltd, 

Chichester, 2002, pp. 165. 

2. I. Marin-Montesinos, D. H. Brouwer, G. Antonioli, W. C. Lai, A. 

Brinkmann, M. H. Levitt, J. Magn. Reson. 177, 307 (2005). 

3. E.H. Hardy, R. Verel and B.H. Meier, J.Mag.Reson.148, 459 (2001). 

4. I. Scholz, B. H. Meier, M. Ernst, J. Chem. Phys. 127, 204504 (2007). 


Sn26 

Solid state NMR RELOADed: increased sensitivity for membrane 

protein studies 

Lopez, Jakob J.; Glaubitz, Clemens 

Goethe University, Frankfurt, Germany 

A notable drawback in NMR is its inherent low sensitivity. 80% of the 

instrument time spent on experiments consists solely of relaxation 

periods during which nuclei regain their Boltzman equilibrium. In the 

study of membrane proteins by solid state NMR, this bottleneck is 

narrowed further by the sample limitations inherent in membrane 

protein studies, which lead to very low sample quantities in the solid 

state MAS rotor. In this conference contribution, 13 C solid state NMR 

experiments are presented which allow the user to acquire spectra 

in time periods which are notably shorter than previously necessary. 

These experiments are band selective in nature and utilize the cooling 

potential of unperturbed nuclei to lower the spin temperature of their 

excited neighbors. As we demonstrate, it becomes possible to replace 

the recycle delay in a series of scans by a time period during which 

proton driven spin diffusion drives the relaxation enhancement by 

lower adjacent spin temperature (RELOAD). A duration of ~200 ms 

suffices for this step, and for 1D 13 C NMR experiments, the omission 

of recycle delays (typically of 2 s length) reduces the measuring 

time by approximately a factor of four. For two dimensional 13 C NMR 

experiments, the measuring time is reduced even further (~8), as the 

band selectivity permits a lower number of increments to be acquired. 

Indeed, for RELOADed 3D solid state NMR experiments, the amount of 

time saved may be envisioned to become dramatic. In this contribution, 

we first investigate the RELOAD effect using the uniformly 13 C labelled 

tripeptide formyl-MLF in the microcrystalline form. The effect of the 

RELOAD delay, the time which the excited nuclei are exposed to their 

cooling reservoir, as well as the RELOAD number, the number of times 

which the cooling reservoir may be called upon before losing its cooling 

ability, is assessed. In the last part of this contribution, a number of 

aspects and first results of RELOADed versions of 2D correlation 

spectra of membrane proteins are presented. 

ABSTRACTS POSTER


Sn27 

95 Mo solid-state NMR study of transition metal cluster 

compounds: A synergetic experimental & computational 

approach 

Cuny, Jerome 1 ; Gautier, Regis 1 ; Le Polles, Laurent 1 ; Trebosc, Julien 2 ; 

Delevoye, Laurent 2 ; Pickard, Chris J. 3 

1 Sciences Chimiques de Rennes, UMR CNRS, Ecole Nationale 

Superieure de Chimie de Rennes, Rennes, France; 2 Université Lille, 

Unité de Catalyse et Chimie du Solide, Lille, France; 3 University College, 

Department of Physics & Astronomy, London, United Kingdom 

Transition metal clusters are chemical units that contain three or more 

metal centers held together, at least partially, by metal–metal bonds. 

From structural and electronic points of view, these compounds stand 

on the threshold between molecular chemistry and bulk chemistry. 

Some of them exhibit interesting physical properties and potential 

applications - e.g., superconductivity with high critical field (60 T), 

thermoelectricity, catalytic or redox intercalation properties [1]. Among 

the transition elements forming clusters, molybdenum is one of the 

most popular in solid-state chemistry. Numerous oxides, chalcogenides, 

and halogenides compounds presenting Mo clusters of diverse 

nuclearities and geometries have already been synthesized [1]. 

During the last few years, jointly to the emergence of nanosciences, 

a new field of research has been developed for the elaboration of 

transition metal cluster materials by soft chemistry route. Since these 

materials are often poorly crystalline, solid-state NMR can provide 

relevant information about their local structure. 95 Mo NMR investigations 

of these hybrid nanomaterials will enable to learn more about their 

chemical interactions with the matrix. In a first step, crystallized 

precursors must be studied. 

We present 95 Mo solid-state NMR analyses of some dimeric units 

as well as trimers, tetramers, and larger molybdenum clusters. To 

overcome difficulties due the low NMR sensitivity of the 95 Mo nucleus, 

a high-field spectrometer (18.6 T) and sensitivity enhancement 

techniques (HS-QCPMG) were employed. To our knowledge, no 

experimental work dealing with 95 Mo solid-state NMR on insulator 

cluster compounds have been reported in the literature. Since the 

complexity of the experimental results do require a theoretical analysis 

for a complete understanding, quantum chemical calculations of 

chemical shift and quadrupolar interaction parameters have been 

performed using DFT calculations based on the GIPAW formalism [2]. 

[1] Clusters and Colloids, ed. G. Schmid, 1994, VCH, Weinheim. 

[2] C. J. Pickard, F. Mauri Phys. Rev. B 2001, 63, 245101. 

104 

Sn28 

TSAR based methods for protein structure determination 

Lewandowski, Józef R. 1 ; De Paëpe, Gaël 2 ; Eddy, Matthew T. 3 ; Lelli, 

Moreno 4 ; Loquet, Antoine 5 ; Bhaumik, Anusarka 4 ; Struppe, Jochem 6 ; 

Maas, Werner 6 ; Böckmann, Anja 5 ; Luchinat, Claudio 7 ; Bertini, Ivano 8 ; 

Griffin, Robert G. 3 

1 Massachusetts Institute of Technology/Université de Lyon, CNRS 

/ ENS-Lyon / UCB-Lyon 1, Centre de RMN à Très Hauts Champs, 

Cambridge/Lyon, United States; 2 Massachusetts Institute of Technology/ 

INAC/SCIB/LRM, CEA Grenoble, Cambridge/Grenoble, United States; 

3 Massachusetts Institute of Technology, Francis Bitter Magnet Lab, 

Cambridge, United States; 4 University of Florence, Magnetic Resonance 

Center, CERM, Sesto Fiorentino, Italy; 5 Institut de Biologie et Chimie 

des Protéines, C.N.R.S UMR 5086, Lyon, France; 6 Bruker BioSpin 

Corporation, Billerica, United States; 7 University of Florence, Magnetic 

Resonance Center, CERM, Sesto Fiorentino/Florence, Italy; 8 University of 

Florence, CERM/Department of Chemistry, Sesto Fiorentino, Italy 

Development of efficient dipolar recoupling magic angle spinning 

experiments that yield long-distance restraints is a crucial component 

necessary for solid-state NMR to become a routine tool for 

macromolecular structure determination. In that spirit, we have recently 

introduced a three-spin second order recoupling mechanism, Third 

Spin Assisted Recoupling (TSAR)[1], that gave rise to a number of 

efficient and highly flexible experiments, including homonuclear Proton 

Assisted Recoupling (PAR)[2,3] and Proton Assisted Insensitive Nuclei 

Cross-Polarization (PAIN-CP)[1]. These methods have yielded numerous 

long-distance 13 C- 13 C, 15 N- 15 N and 15 N- 13 C restraints in several protein 

systems. 

In this contribution we give an overview of the principles governing 

TSAR recoupling highlighting its advantages for application at high 

magnetic fields and high spinning frequencies – an attractive regime for 

macromolecular structural studies for which sensitivity and resolution 

provide the most stringent limitations. We place TSAR in the context 

of protein structure determination, including a number of examples 

from 15 N- 15 N spectroscopy and applications of PAR for obtaining longdistance 

13 C- 13 C contacts in proteins at very high spinning frequencies 

(up to 65 kHz) to high-resolution structure determination of the Crh 

protein dimer (86 AA 2x10.4 kDa)[4] and the catalytic domain of 

the matrix metalloproteinase MMP-12 (159 AA, 17.6 kDa)[5]. We 

discuss the TSAR based experiments as valuable tools on their own 

and powerful complementary techniques to the other solid-state NMR 

methods. 

[1] Lewandowski, J. R.; De Paepe, G.; Griffin, R. G. J. Am. Chem. Soc. 

2007, 129, 728-729. 

[2] De Paepe, G.; Lewandowski, J. R.; Loquet, A.; Bockmann, A.; Griffin, 

R. G. J. Chem. Phys. 2008, 129, 245101: 1-21. 

[3] Lewandowski, J. R.; De Paepe, G.; Eddy, M. T.; Griffin, R. G. J. Am. 

Chem. Soc. 2009 DOI:10.1021/ja806578y. 

[4] Bockmann, A.; Lange, A.; Galinier, A.; Luca, S.; Giraud, N.; Juy, M.; 

Heise, H.; Montserret, R.; Penin, F.; Baldus, M. J Biomol NMR 2003, 27, 

323-39. 

[5] Balayssac, S.; Bertini, I.; Bhaumik, A.; Lelli, M.; Luchinat, C. Proc. 

Natl. Acad. Sci. U. S. A. 2008, 105, 17284-9. 


Sn29 

Isotropic resolution for 3/2 spin quadrupolar nuclei by inverse 

detection of spin ½ 

Trébosc, Julien; Wang, Qiang; Hu, Bingwen; Amoureux, Jean-Paul 

University of Lille, UCCS, Villeneuve d’Ascq, France 

Heteronuclear correlation spectra involving quadrupolar nuclei with 

isotropic resolution are relying on ‘single nucleus’ quantum experiments 

usually MQ or STMAS sequence followed by magnetization transfer 

through CP, J or dipolar INEPT mechanism [1,2]. 

Here we present a new dipolar-driven HMQC sequence using a t - 1 

split-STMAS [3] block in the indirect dimension to obtain isotropic 

resolution for spin 3/2. We call this sequence D-HMQC-ST. Dipolardriven 

D-HMQC (or D-HSQC) methods use either SFAM or symmetry 

1 based SR4 heteronuclear dipolar recoupling techniques. On the 

2 

indirect channel, a fully symmetric version of t -split STMAS allows a 

1 

proper reconversion of multiple-nucleus multiple quantum coherences 

back into single-quantum observable coherences. We achieve transfer 

efficiencies of the order of 20-30% for D-HMQC-ST, as compared to 

that obtained with a central transition selective 90° pulse. This makes 

the concept very attractive for producing isotropic quadrupolar spectra 

in the indirect dimension. This lower efficiency is counter-balanced by 

much narrower linewidth and thus overall S/N is often improved using 

D-HMQC-ST, as compared to a ‘classical’ D-HMQC method. 

The experimental proofs of this concept are shown on 1H-23Na as well 

as 31P-23Na D-HETCOR spectra of sodium-hydrogeno-phosphate and 

sodium di-hydrogeno-phosphate samples on 4, 3.2 and 1.3mm MAS 

probes at 10, 20 and 60 kHz MAS speed. For 31P observed D-HMQC-ST 

spectra we start from a preliminary 1H-31P CP because of longer 31P longitudinal relaxation times. 

This technique opens a very interesting new way to do inverse detection 

of spin-3/2 low gamma quadrupolar nuclei with isotropic resolution as 

well as quadrupolar to quadrupolar D-HETCOR spectra with potentially 

isotropic resolution in both dimensions. 

1- Wang, S. H.; De Paul, S. M. and Bull, L. M. ; J. Magn. Res. 125 

(1997) 364 

2- Wiench, J. W. & Pruski, M. ; Solid State Nucl. Magn. Reson. 26 

(2004) 51 

3- Trébosc, J.; Amoureux, J. & Gan, Z. ; Solid State Nucl. Magn. Reson. 

31 (2007) 1 

Sn30 

Weathering and crystallisation of a phosphate glass studied 

through 17 O solid-state NMR and first-principles calculations 

Forler, Nina 1 ; Delevoye, Laurent 1 ; Vasconcelos, Filipe 1 ; Cristol, Sylvain 1 ; 

Paul, Jean-François 1 ; Mauri, Francesco 2 ; Charpentier, Thibault 3 ; 

Montagne, Lionel 1 

1 UCCS Université de Lille - Nord de France, Villeneuve d’Ascq, France; 

2 Université Pierre et Marie Curie, Institut de Minéralogie et Physique des 

Milieux Co, Paris, France; 3 CEA, IRAMIS, Service de Chimie Moléculaire, 

Gif sur Yvette, France 

Phosphate-based compounds are a very important class of materials in 

various chemistry domains that they are involved in (glasses, ceramics, 

gels, biomaterials, catalytic precursors, ionic conductors, laser 

materials, water-softening agents,…). If 31 P is largely used for structural 

characterisation of crystalline and amorphous phosphates through 

Solid-State MAS NMR, very few 17 O MAS NMR studies have been 

reported in the literature due to 1) low natural abundance of 17 O isotope, 

2) low sensitivity and 3) second-order quadrupolar broadening. Even 

fewer studies were conducted on hydroxylated phosphate compounds, 

due to the complexity of obtaining the corresponding 17 O-enriched 

samples. However, there is a large interest for the study of hydroxylated 

phosphate compounds since they are involved in many processes 

and applications, e.g. baking powder, glass precursors, alteration 

products of biomaterials. In the present study, we used a combination 

of 17 O NMR methods at a very high magnetic field with first-principles 

calculations in order to characterize the oxygen sites in a series of 

hydroxylated sodium phosphate compounds. The determination of their 

chemical shifts and quadrupolar parameters were interpreted in terms 

of local and semi-local environment, [1] i.e. the nature of the cationic 

surrounding, the covalence of the bonds as well as the nature of the 

hydrogen bonding. The magnitude of the quadrupolar interaction and 

its asymmetry revealed to be a precise indicator of the local structure 

in sodium hydrogen phosphates. The calculations, performed using the 

GIPAW formalism, developed by Pickard and Mauri [2], gave access 

to the NMR parameters, for a large series of phosphate structures, 

hence many of them did not need to be synthesized. Our 17 O NMR 

experimental and computing approach allowed to accurately describe 

the weathering mechanism of a phosphate glass, including identification 

and quantification of the different crystalline phases that are formed 

during the alteration process. 

[1] Vasconcelos, F., et al., Inorganic Chemistry, 2008, 47, 7327-7337 

[2] Pickard, C. J. & Mauri, F. Phys. Rev. B, 2001, 63, 245101 


Sn31 

Membrane interactions of Bcl-2 proteins involved in apoptosis 

Wallgren, Marcus 1 ; Pedersen, Anders 2 ; Sani, Marc-Antoine 1 ; Dufourc, 

Erick J. 3 ; Gröbner, Gerhard 1 

1 Umeå University, Department of Chemistry, Umeå, Sweden; 2 Göteborg 

University, Swedish NMR Centre, Göteborg, Sweden; 3 2CNRS-Université 

Bordeaux, Bordeaux, France 

Escape from programmed cell death, apoptosis, is one of the main 

hallmarks of cancer. The anti-apoptotic protein Bcl-2 belongs to the 

Bcl-2 protein family, which function as a major gatekeeper in the 

mitochondrial apoptotic pathway. Bcl-2 is found to a great extent in 

many breast cancers and is highly involved in the inherent resistance to 

anti-cancer drugs. This protein is mitochondrial membrane-associated 

and we will use NMR spectroscopy to provide structural information of 

the membrane-mediated mechanism underlying the action of Bcl-2 as a 

potent inhibitor of cell death. For this purpose we currently work on the 

expression of the full-length Bcl-2, both in vivo and in vitro. In addition, 

the full-length pro-apoptotic Bax protein, the counterplayer of Bcl-2, is 

also expressed. Bax is upon activation translocated to the mitochondrial 

membrane where it forms oligomers, leading to pore formation, release 

of cytochrome c and cell death. Studying the interplay between Bax 

and Bcl-2 when both are present at the membrane level will reveal 

information important for understanding the cell protecting mechanism 

of Bcl-2. Preliminary studies of the membrane dependent behaviour 

of synthesized segments of these two proteins have been carried out 

using a number of biophysical methods ranging from CD (Circular 

Dichroism), ATR (Attenuated Total Reflection), Calorimetry to solid 

state NMR spectroscopy. Our first results reveal that the unique BH4 

domain of the Bcl-2 protein requires cardiolipin to be able to convert 

into an á-helix conformation, enabling membrane-protein interaction 

which seems essential to block any apoptotic action with its connected 

pore-formation and cytochrome c release. In this process the first helix 

localized at the N-terminus of Bax (Bax-á1) can act as an addressing 

sequence, which upon activation directs the protein from the cytosol 

towards the mitochondria. 

ABSTRACTS POSTER


Sn32 

Methodology of solid state NMR study of reversible guest 

exchange in porous materials 

Gajda, Jaros³aw; Jeziorna, Agata; Potrzebowski, Marek 

Polish Academy of Sciences, Lodz, Poland 

Porous materials (PM) have recently received the great deal of interest 

due to their prospective practical applications. Such compounds 

can be useful in gas storage, as molecular sieves, size- and shapeselective 

catalysis, magnetism, optoelectronic devices. Last decade 

has witnessed the explosive growth in methods of synthesis and 

characterization of PM. Understanding and controlling the topological 

and geometric relationships between host-guest, along with the 

characteristics of the host matrices is key step in producing of new 

materials with desired properties. 

Number of experimental techniques can be used in structural studies 

of PM. The most important are diffraction methods providing the 

comprehensive and in many cases unequivocal set of structural data. 

Unfortunately, a lot of PM materials are not amenable to a structure 

determination based on single-crystal diffraction methods (neutron/Xray). 

This can be caused by a variety of reasons such as the inability 

to grow large single crystals, inherent disorder within a crystal, or 

defects caused by impurities. Phenomena such as phase transitions, 

polymorphism, sorption/desorption processes have significant influence 

on structure of materials. Solid state NMR (SS NMR) spectroscopy is an 

unique technique which allows investigating given supra phenomena 

with great accuracy. SS NMR is a “fingerprint” of local structure and 

represents local electronic environment for each nuclei. 

In this work we wish to present power of high-resolution 1D NMR (BD, 

CP/MAS) and 2D NMR (2D PASS, FSLG HETCOR, POST-C7) solid state 

spectroscopy in structural studies of porous materials with organic, 

hetero-organic and metal-organic frameworks. The new results showing 

influence of reversible migration of guest molecules (organic solvents 

and/or water) on geometry of host lattice will be discussed. SS NMR 

results will be compared with X-ray and theoretical data obtained by 

using DFT GIAO calculations. 

Sn33 

NMR evidence for molecular groups trapped in cages of Co 4 Al 13 , 

Fe 4 Al 13 and Ru 4 Al 13 

Bobnar, Matej 1 ; Vrtnik, Stanislav 1 ; Jegliè, Peter 1 ; Klanjšek, Martin 1 ; 

Dolinšek, Janez 1 ; Bauer, Birgitta 2 ; Gille, Peter 2 ; Heggen, Marc 3 ; 

Feuerbacher, Michael 3 ; Haarmann, Frank 4 ; Burkhardt, Ulrich 4 ; Grin, Yuri 4 

1 Jožef Stefan Institute, Ljubljana, Slovenia; 2 Ludwig-Maximilians- 

Universität München, München, Germany; 3 Institut für 

Festkörperforschung, Forschungszentrum Jülich, Jülich, Germany; 

4 Max-Planck-Institut für Chemische Physik fester Stoffe, Dresden, 

Germany 

Orthorhombic Co 4 Al 13 with layered structure and pentagonal channels 

[1] has been for a long time considered as an approximant for 

decagonal quasicrystals. Moreover, with 102 atoms per unit cell, cluster 

arrangement of atoms and inherent disorder [1], it has been treated as 

a member of complex metallic alloys [2]. Recent X-ray investigation on 

high-quality single-crystals in combination with theoretical calculations 

[3] revealed its double nature, covalent and ionic, relating them to 

the crystal structures of intermetallic clathrates (cage compounds). 

Co-Al-Co molecular groups play the role of the guest ions trapped in 

cages formed by other Al and Co atoms. Here we present [4] a clear 

evidence for clathrate-like structure of Co 4 Al 13 , based on 27 Al nuclear 

magnetic resonance experiments, which already proved to give insight 

into the atomic structure of similar intermetallic compounds [5,6]. 

Two 27 Al signals were identified in Co 4 Al 13 . First one originates from Al 

atoms forming cages. The second signal corresponds to Al sites with 

106 

exceptionally large almost axially symmetric quadrupole coupling, in 

perfect agreement with isolated Co-Al-Co molecular groups. Both 27 Al 

signals also exhibit different spin-lattice relaxation time T 1 . Similar 

structural features were found in quasihomologous monoclinic Fe 4 Al 13 

[7] and Ru 4 Al 13 . 

References: 

[1] Yu. Grin, U. Burkhardt, M. Ellner, K. Peters, J. Alloys Compd. 206, 

243 (1994). 

[2] K. Urban, M. Feuerbacher, J. Non-Cryst. Sol. 334&335 , 143 (2004). 

[3] Yu. Grin, B. Bauer, U. Burkhardt, R. Cardoso-Gil, J. Dolinšek, M. 

Feuerbacher, P. Gille, F. Haarmann, M. Heggen, P. Jegliè, M. Müller, S. 

Paschen, W. Schnelle, S. Vrtnik. EUROMAT 2007: European Congress on 

Advanced Materials and Processes, Nürnberg, Germany, 2007. 

[4] P. Jegliè, M. Heggen, M. Feuerbacher, B. Bauer, P. Gille, and F. 

Haarmann, J. Alloys Compd. (in press). 

[5] P. Jegliè, M. Komelj, M. Klanjšek, U. Tkalec, S. Vrtnik, M. 

Feuerbacher, and J. Dolinšek, Phys. Rev. B 75, 014202 (2007). 

[6] F. Haarmann, M. Armbrüster, Yu. Grin, Chem. Mater. 19, 1147 

(2007). 

[7] Yu. Grin, U. Burkhardt, M. Ellner, K. Peters, Z. Kristallogr. 209, 479 

(1994). 

Sn34 

Homonuclear dipolar recoupling of quadrupolar spins: New 

developments and applications 

Eden, Mattias; Lo, Andy Y H 

Stockholm University, Physical Chemistry Division, Stockholm, Sweden 

As opposed to the case of spin-1/2 nuclei in solids, relatively few 

options exist for efficiently recoupling the through-space dipolar 

interactions between homonuclear half-integer quadrupolar spins 

undergoing magic-angle spinning (MAS). This partly stems from the 

problems associated with controlling the dynamics of half-integer 

quadrupolar spins and the necessity to employ weak rf fields for 

recoupling. 

We will present our recent advances in this area, including the 

introduction of new symmetry-based dipolar recoupling pulse 

sequences for efficient excitation of double-quantum (2Q) coherences 

between the central transitions of half-integer quadrupolar spins 

in crystalline as well as amorphous inorganic materials. Several 

applications will be demonstrated, for example (1) the possibility to 

determine absolute orientations of 23 Na efg tensors in crystalline 

materials by 2Q NMR; (2) the use of 27 Al 2Q-1Q correlation experiments 

for connectivity-mapping of aluminum polyhedra in aluminate and 

alumino-silicate glasses. 

Sn35 

Exploring Solid-State 17 O NMR to Distinguish Secondary 

Structures in Alzheimer’s Aβ Fibrils 

Antzutkin, Oleg N. 1 ; Filippov, Andrei 2 ; Wong, Alan 3 ; Baldus, Johanna 3 ; 

Hung, Ivan 3 ; Kukol, A. 4 ; Brown, Steven P. 3 ; Smith, Mark E. 3 ; Dupree, Ray 3 

1 University of Warwick and LuleŒ University of Technology, Physics, 

Coventry, United Kingdom; 2 LuleŒ University of Technology, Chemical 

Engineering, LuleŒ, Sweden; 3 University of Warwick, Physics, Coventry, 

United Kingdom; 4 University of Warwick, Chemistry, Coventry, United 

Kingdom 

A vast number of studies has revealed that Alzheimer’s amyloidβ-peptide 

deposits contribute directly to the disease’s progressive 

neurodegeneration. Aggregation cascade for Aβ peptides and structure 

of amyloid fibrils have been previously discussed [1]. Different types of 

Aβ amyloid fibrils have different degree of neurotoxicity in vitro [2]. Aβ 

oligomers are also neurotoxic in vitro since the molecular chaperone 


αB-crystallin prevents Aβ from forming amyloid fibrils but nevertheless 

enhances Aβ toxicity [3]. Antibodies for Aβ oligomers prepared by 

nanogold technology decrease toxicity of Aβ for SH-SY5Y human 

neuroblastoma cell cultures in vitro [4]. Thus, studies on structure of 

Aβ-oligomers/fibrils and interaction of Aβ with biomembranes is of an 

importance [5]. 

We explore solid-state 17 O NMR on selectively 17 O, 13 C, 15 N-labeled 

Aβ(1-40), Aβ(11-25) and Ac-Aβ(16-22)- )-NH 2 peptides to distinguish a 

parallel and anti-parallel β-sheet secondary structures in amyloid fibrils. 

Aβ(1-40) fibrils form in-registry parallel β-sheets [6], while Aβ(11-25) 

[7] and Ac-Aβ(16-22)- )-NH 2 [8] form different anti-parallel β-sheet 

structures, which were previously identified by 13 C multiple-quantum 

and 13 C{ 15 N} REDOR ss-NMR. We found that 17 O NMR chemical shifts 

are sensitive to the type of the secondary structure, i. e. a parallel vs. 

an anti-parallel β-sheet structures, while the quadrupolar parameters 

of 17 O nuclei do not vary beyond the error limits in the simulated 

lineshapes of both fibrillized and unfibrillized peptide systems. Results 

of ss-NMR techniques to measure heteronuclear distances, 15 N{ 17 O}- 

REAPDOR, 15 N{ 17 O}-TRAPDOR and 17 O{15N}-REDOR on selectively 

17O-Val18 and 15 N-Phe20 labeled Ac-Aβ(16-22)-NH 2 fibrils are also 

discussed. These novel ss-NMR experiments are additional tools for 

measuring hydrogen bonding of peptides in amyloid fibrils. 

[1.] Antzutkin MRC 42 (2004) 231 

[2.] Petkova et al Science 307 (2005) 262 

[3.] Stege et al BBRC 262 (1999) 152 

[4.] Kayed et al Science 300 (2003) 486 

[5.] Bokvist et al JMB 335 (2004) 1039 

[6.] Antzutkin et al PNAS 97 (2000) 13045 

[7.] Petkova et al JMB 335 (2004) 247 

[8.] Balbach, Ishii, Antzutkin et al Biochemistry 39 (2000) 13748. 

Sn36 

NMR study of magnetic field distortions in ferroelectric liquid 

crystals 

Dong, Ronald 

University of British Columbia, Vancouver, Canada 

Ferroelectric liquid crystals (FLCs) have attracted much interest due to 

their applications in electro-optical devices. Deuterium nuclear magnetic 

resonance (DNMR) has been used in the past to study soliton-like 

distortions by observing the spectral line shape as a function of the 

NMR field direction in aligned deuteriated FLC samples with respect to 

their helical axes. The powerful technique of angle dependent DNMR 

spectroscopy in FLCs will be highlighted. In chiral smectic C* phases, an 

in-plane electric polarization exists spontaneously in each smectic layer. 

When the helix is not distorted, the modulation wave along the pitch 

axis has a linear phase profile. The helicoidal structure may, however, 

be distorted in the presence of an external magnetic field. Domains are 

then formed to give a soliton-like lattice where the molecules, mostly 

aligned by the field, are separated by narrow domain walls in which 

the molecules twist rapidly. As a result, a non-linear spatial phase 

modulation profile is seen. If the applied field is large enough, the 

helices can even be unwound to give a homogeneous smectic C phase. 

Spectral simulation based on the Landau theory is used to assess the 

critical magnetic field strength for unwinding the helices. In one FLC, 

the winding/unwinding transitions are observed at different magnetic 

field strengths when the field direction is along the pitch. In other FLCs, 

the critical field has been determined at different temperatures far away 

from the Lifshitz point. The structure and degree of distortions can 

also be monitored from the spectral simulations. These results will be 

discussed and contrasted. 


Sn37 

Real time 31 P and steady state 31 P/ 13 C solid state NMR on the 

integral membrane protein diacylglycerol kinase from E.coli 

Wollschlag, Sandra Johanna; Hellmich, Ute Andrea; Glaubitz, Clemens 

Goethe Universität Frankfurt, Institute for Biophysical Chemistry, BMRZ, 

Frankfurt a.M., Germany 

The integral membrane protein Diacylglycerol Kinase (DGK) from E.coli 

is used as a model system for structure-function studies on a lipid 

regulator in its native environment, the lipid bilayer. The DGK monomer 

consists of three transmembrane helices and two amphiphatic helices 

with orientation to the cytoplasm. In its active conformation, DGK forms 

a homotrimer with one putative active site per monomer. DGK catalyses 

the transfer of the γ-phosphate of Mg*ATP to diacylglycerol to generate 

phosphatidyl acid at the interface membrane-cytoplasm. Both, substrate 

and product are important signalling molecules. 

Solid-state NMR is a unique tool for the investigation of membrane 

proteins in their native environment and for probing enzymatic reactions 

regardless of their compartmentalization. For such experiments, the 

amount of protein and the quality of the sample preparation are crucial. 

Expression, purification, reconstitution and sample preparation were 

optimised, so that a sample of DGK in high quality became available 

in amounts necessary for ssNMR experiments, while maintaining its 

specific activity. 

A 31 P Real Time MAS experiment was designed and implemented, 

to investigate for the first time ATP hydrolysis and substrate 

phosphorylation by a membrane protein inside the lipid bilayer 

simultaneously and with atomic resolution. This experiment yields rate 

constants and stoichiometry. Furthermore, inhibition experiments were 

carried out. Now it is possible to characterise the enzymatic mechanism. 

This experiment also shows, that membrane proteins can be kept fully 

active under the conditions of Magic Angle Sample Spinning, which 

involves high spinning speeds (hence centrifugal forces), high protein 

concentrations, reduced hydration and low lipid:protein ratios. 

Concepts to extend this experimental approach towards studying 

conformational changes during the enzymatic cycle within DGK will also 

be presented and discussed. The aim is a detailed understanding of the 

interactions of an integrale membrane protein with its substrates as well 

as the surrounding lipids, their influence on conformation, structure, 

dynamic and vice versa by ssNMR. 

Sn38 

Investigation of analysis for vulcanized natural rubber by 

inverse measurements with FG-MAS probe 

Sakurai, Satoshi 1 ; Utsumi, Hiroaki 1 ; Kawahara, Seiichi 2 

1 JEOL Ltd., Application & Research group, Tokyo, Japan; 2 Nagaoka 

University of Technology, Department of Chemistry, Nagaoka, Japan 

Analysis of crosslinking junctions of crosslinked rubber samples 

has been made through solid-, solution- and latex-state NMR 

spectroscopies up to date. If we apply ordinary pulse techniques 

of solution-state NMR spectroscopy under MAS, it is possible to 

positively analyze the crosslinking junctions, more exactly, because the 

rubber samples have high mobility. In this case, the analysis through 

inverse measurements are quite effective to shorten a time for the 

measurement. However, because the signals assigned to the main 

chains are too big, the huge correlation signals of the main peaks 

interfere with the small signals of the crosslinking junctions under a 

resolution of the 1H observation. In the present study, an attempt to 

analyze the crosslinking junctions of the rubber samples was made by 

solution-state NMR spectroscopy under MAS with selective neglection 

of the main correlation signals. 

ABSTRACTS POSTER


Sn39 

Pulsed RF µSR and composite pulses 

Clayden, Nigel 1 ; Cottrell, Stephen 2 ; McKenzie, Iain 2 

1 University of East Anglia, Chemical Sciences and Pharmacy, Norwich, 

United Kingdom; 2 STFC, ISIS Facility, Didcot, United Kingdom 

Complementary to conventional magnetic resonance techniques, muon 

spectroscopy [1] uses an implanted 100% spin polarised spin-½ 

positive muon either as a probe of the local magnetic environment or to 

act as a proton analogue in diffusion studies and chemistry. 

With due regard to the short muon lifetime (~2.2µs), modern pulsed RF 

techniques can be applied; however, early work [2, 3] has demonstrated 

that the potential for finite pulse artefacts is significant. These arise 

because firstly, the large magnetogyric ratio (~13.6kHz/G) leads to 

strong dipolar couplings and large resonance offsets, and secondly, the 

RF coils are generally large and poorly shaped (~24x24x2mm) to match 

the profile of the incoming muon beam and give poor RF homogeneity. 

Recently, we have explored the application of composite pulses as 

a means of improving the RF excitation profile. The work presented 

demonstrates a 90 x 180 y 90 x sequence as a means of providing spin 

inversion [4], and compares its performance in the context of the 

RF µSR experiment to that obtained from using simple 180° pulses. 

Intriguingly, because detection is made by monitoring muon decay 

products, the effect of pulses on the muon can be observed throughout 

the composite sequence, raising the possibility of using RF µSR as a 

tool for studying spin evolution during RF sequences. 

References 

[1] S.F.J. Cox, J.Phys. C: Solid State Phys. 20 (1987) 3187 

[2] S.P. Cottrell et al, Appl.Magn.Reson. 15 (1998) 469. 

[3] N.J. Clayden et al, Phys.Chem.Chem.Phys. 8 (2006) 3094. 

[4] M.H. Levitt and R. Freeman, J.Magn.Reson. 33, 473 (1979). 

Sn40 

Solid-state NMR studies of polymer nanocomposites 

Vyalikh, Anastasia 1 ; Massiot, Dominique 2 ; Scheler, Ulrich 1 

1 Leibniz Institute of Polymer Research Dresden, Dresden, Germany; 

2 CEMHTI UPR3079 CNRS, Université d’Orléans, Orleans, France 

Incorporating inorganic fillers into polymers mechanical and rheological 

properties of composite materials are significantly improoved. Nanosized 

fillers have the advantage of a large of nanofiller–matrix interface. 

One challenge is to achieve a high degree dispersion of nanoparticles 

throughout the polymer matrix. In this study layered double hydroxides 

(LDH), a synthetic clay with layered structure, has been used [1]. 

Compatibility with a non-polar polymer matrix and expansion of 

interlayer space in LDH are achieved by intercalation of surfactant 

molecules [2] 27Al MAS and 27Al triple-quantum (3Q)-MAS NMR 

has been applied to monitor and quantify the structural changes as a 

result of surfactant modification of LDH .[3] Two-dimensional one pulse 

spectrum (TOP) [4] representation of the MAS spectra facilitates the 

analysis of the satellite transitions. In pure LDH only six-coordinated, 

octahedral, aluminium is found. In calcined LDH treated by a surfactant 

in addition a considerable fraction of four-coordinated, tetrahedral, 

aluminium has been identified. Applying two-dimensional 3QMAS 

NMR, two sites with significantly different quadrupolar parameters are 

resolved, which permit quantification of the components A quantitative 

analysis based on the 27Al MAS spectra reveals that depending on the 

type of surfactant in the modified sample between 24% and 31% of the 

aluminium has been converted to four-fold coordination.[5] Additional 

insight is gained from high-resolution solid-state 1H NMR, which gives 

detailed information of the water and surfactant. Different hydrogen 

bonded states of OH protons are distinguished by proton T1rho 

experiments, resolving different sites. 

108 

References 

[1] F.R. Costa, A. Leuteritz, U. Wagenknecht, D. Jehnichen, L. Häuβler, G. 

Heinrich, Appl. Clay Sci. 38 (2008) 153-164. 

[2] J. Rocha, M. del Arco, V. Rives, and Ulibarri, M.A.,. J. Mater. Chem. 9 

(1999) 2499-2503. 

[3] Sideris J.P, Nielsen, U.J., Ghan, Z., Grey, C.P., Science, 321, p. 113, 

2008 

[4] P. Blümler, J. Jansen, and B. Blümich, , Solid State NMR 3 (1994) 

237-240. 

[5] A. Vyalikh, D. Massiot, U. Scheler, Solid-state NMR (submitted) 

Sn41 

Effect of B/Al substitution on crystallization and structure of 

boron gehlenite: A combined Neutron Powder Diffraction and 

High Resolution NMR study 

Montouillout, Valerie; Véron, Emmanuel; Florian, Pierre; Massiot, 

Dominique; Allix, Mathieu; Ory, Sandra; Matzen, Guy 

CEMHTI-CNRS, Orléans, France 

The aluminium substitution by boron into alumino-silicate glassy or 

crystalline materials induced significant modifications of chemical 

properties of great interest for industries (viscosity, melting and 

crystallisation temperatures,..). 

The aim of the present work is to study the incorporation of boron, in 

substitution for aluminium in the structure of gehlenite Ca2Al2SiO7, 

mineral of the melilite family. This network presents two non equivalent 

tetrahedral sites for aluminium and silicon: T1 exclusively occupied 

by aluminium, and T2 occupied either by aluminium or by silicon. The 

questions are: where does the boron be included, and what are the 

consequences for the structure? 

Samples with various amount of boron were crystallized from the 

Ca2Al2-xBxSiO7 (with 0 ≤ x ≤ 2) glass. A structural analyses by neutron 

powder diffraction was completed by a precise description of 11B, 27Al 

and 29Si environments using high resolution solid-state NMR. 

In a first step, we propose the assignment of 29Si and 27Al MAS and 

MQ-MAS signals representative respectively of the two silicon sites and 

the seven aluminum sites of Ca2Al2SiO7. This assignment is based 

on homonuclear (INADEQUATE) and heteronuclear (INEPT) J coupling 

correlations experiments and allows the complete description of the 

network. 

In a second step, the effects of boron substitution were studied by 

Rietveld refinements of X-ray and neutron powder diffraction data and 

multiple resonances solid state NMR experiments. The localization of 

boron atoms and the local distortions induced are discussed. 

Sn42 

1 H-driven 19 F spin diffusion and cross polarization 

measurements for distance and structure determination in 

ciprofloxacin derivatives 

Garro Linck, Yamila 1 ; Chattah, Ana Karina 1 ; Romañuk, Carolina 2 ; Olivera, 

María Eugenia 2 ; Manzo, Rubén H. 2 ; Cuffini, Silvia 3 ; Raya, Jesus 4 ; 

Hirschinger, Jerome 4 ; Monti, Gustavo A. 1 

1 Facultad de Matemática, Astronomía y Física, Universidad Nacional 

de Córdoba, Córdoba, Argentina; 2 Facultad de Ciencias Químicas, 

Universidad Nacional de Córdoba, Córdoba, Argentina; 3 CEPROCOR, 

Ministerio de Ciencia y Tecnología, Córdoba, Argentina; 4 Institute Le Bel 

de la Universidad Louis Pasteur, Strasbourg, France 

Dipolar-driven spin diffusion in solid state Nuclear Magnetic Resonance 

(NMR) is a robust approach for obtaining homonuclear distances. In this 

work, we applied the centerband-only detection of exchange (CODEX) 

technique [Wenbin Luo and Mei Hong. J. Am. Chem. Soc. 2006, 

128, 7242-7251] on a new pharmaceutical compound, ciprofloxacin 


saccharinate [Romañuk et al. J. Pharm. Sci. DOI: 10.1002/jps.2168]. 

This compound exhibits polymorphism, and presents well distinguished 

solid forms, CIP-SAC (I) and CIP-SAC (II). In particular, CIP-SAC (II) can 

be obtained as a monocrystal. 

To carry out the analysis, we use crystallographic data obtained from 

single crystal X-ray diffraction of CIP-SAC (II), as a starting point to 

perform the fittings of CODEX data. We calculate the value of F (0), ij 

the overlap integral describing the probability that single-quantum 

transitions occur at the same frequency for spins i and j, for CIP-SAC 

(II). Then, considering this value, we were able to determine distances 

and molecules per unit cell in CIP-SAC (I). 

Diffraction data show that CIP-SAC (II) presents two molecules per unit 

cell (one in the asymmetric unit) with different orientations. From 13C NMR data, CIP-SAC (I) shows at least two molecules per asymmetric 

unit. Results from CODEX, in addition to 19 F 13 F spectra, T and C CP-MAS 

1 

(cross polarization and magic angle spinning) experiments performed 

on both solid forms of CIP-SAC, give us evidence that CIP-SAC (I) is a 

mixture of two polymorphs, one of them probably CIP-SAC (II). 

We complemented these experiments with 1H-19F CP-MAS experiments 

with variable contact time. 

Sn43 

2D homonuclear and heteronuclear correlation NMR 

experiments in solid state applied to a new pharmaceutical 

compound 

Garro Linck, Yamila 1 ; Chattah, Ana Karina 1 ; Romañuk, Carolina 2 ; Olivera, 

María Eugenia 2 ; Manzo, Rubén H. 2 ; Cuffini, Silvia 3 ; Graf, Robert 4 ; Monti, 

Gustavo A. 1 ; Spiess, Hans W. 4 

1 Facultad de Matemática, Astronomía y Física, Universidad Nacional 

de Córdoba, Córdoba, Argentina; 2 Facultad de Ciencias Químicas, 

Universidad Nacional de Córdoba, Córdoba, Argentina; 3 CEPROCOR, 

Ministerio de Ciencia y Tecnología, Córdoba, Argentina; 4 Max-Planck 

Institute for Polymer Research, Mainz, Germany 

Ciprofloxacin (CIP) is a widely prescribed broad-spectrum oral 

fluoroquinolone antibiotic. Low solubility and bitter taste are unfavorable 

properties associated with most fluoroquinolones that limit their 

formulation. Saccharine (SAC) derivatives, either as salts or co-crystals, 

show an improved solubility behavior and better taste. 

Advanced solid state nuclear magnetic resonance (NMR) techniques 

have been used to study a new pharmaceutical compound, the 

ciprofloxacin saccharinate, CIP-SAC. This compound has been 

previously reported and fully characterized with a variety of solid state 

techniques (IR, DSC, XRD and 13 C CP-MAS NMR). [Romañuk et al. 

J. Pharm. Sci. DOI: 10.1002/jps.2168] This saccharinate evidences 

polymorphism presenting two well distinguished solid forms, CIP-SAC 

(I) and (II). 

In this work we performed 2D 13 C- 1 H heteronuclear correlation 

experiments (REPT-HSQC and HETCOR) which allowed the complete 

assignment of the resolved 1 H solid state spectrum. [Kay Saalwächter, 

Robert Graf and Hans W. Spiess. J. Mag. Reson. 

140, 471-476 (1999) and van Rossum et al. J. Mag. Reson. 124, 516– 

519 (1997)] These data were complemented with double quantum MAS 

experiments (BABA pulse sequence [M. Feike, D. E. Demco, R. Graf, J. 

Gottwald, S. Hafner, and H. W. Spiess. J. Mag. Reson. A 122, 214-221 

(1996)]) to identify 1 H- 1 H connectivities, hydrogen bonds and dipolar 

proximities. The zwitterionic character of ciprofloxacin and its derivative 

was identified. Further information about inter and intra molecular 

interactions in both solid forms of the new saccharinate was obtained. 

Differences and similarities in the structures of the different solid 

forms have also been established and supported with single crystal XR 

diffraction data obtained for CIP-SAC (II). 


Sn44 

Phenol-polymer proximity in thermoresponsive gels determined 

by solid state 1H CRAMPS NMR spectroscopy 

Domján, Attila 1 ; Geissler, Erik 2 ; László, Krisztina 3 

1 Chemical Research Center, Hungarian Academy of Sciences, Institute 

of Structural Chemistry, Budapest, Hungary; 2 Université J. Fourier de 

Grenoble, 2Laboratoire de Spectrométrie Physique CNRS UMR558, 

Grenoble, France; 3 Budapest University of Technology and Economics, 

3Department of Physical Chemistry and Materials Sc, Budapest, 

Hungary 

This work reports results obtained by 1H CRAMPS in an amorphous 

soft material. By using an NMR technique (1H-1H correlation CRAMPS) 

that has hitherto been successfully applied mostly to crystalline 

systems of small molecules, we determine approach distances 

between the network chains of an amorphous polymer gel (poly(Nisopropyl-acrylamide)) 

and guest molecules (phenol) above the lower 

critical solution temperature (LCST). This development provides direct 

information on the distribution of guest molecules at the polymer – 

solvent interface that is otherwise difficult to obtain. 

To achieve a two dimensional correlation spectra PMLG-5 sequences 

were used in the indirect dimension and wPMLG-5 sequences in 

the direct dimension. Correlation cross peak volumes measured as 

a function of correlation time were analyzed by rate matrix approach 

introduced by Macura and Ernst. The simplified analysis of the data 

yields an estimate of 5 Å for the distance between the phenol and 

the polymer side-chains. This short distance implies second order 

interactions, most probably an H-bond between the phenol OH group 

and the polymer carbonyl or NH groups. 

Our results show that two-dimensional CRAMPS experiments are not 

limited to crystalline materials, but can also be applied to amorphous 

systems that display only local ordering. From these NMR experiments 

we conclude that the residence time of phenol near the polymer sidechain 

is long. The ability to estimate the phenol – side-chain distance 

holds promise for a variety of amorphous or mobile substrates, such 

as drug delivery vectors or biological systems, and for other dissolved 

molecules than phenol. 

This work was supported by the Hungarian project GVOP-3.2.1.-2004- 

04-0210/3.0 and Hungarian National Research Fund (OTKA, grant No. 

K75182). 

Sn45 

2 H NMR determination of order parameters and phase diagram 

in confined photosensitive liquid crystal 

Zupancic, Blaz 1 ; Zalar, Bostjan 1 ; Diez Berart, Sergio 2 ; Finotello, Daniele 3 ; 

Lavrentovich, Oleg 3 

1 Jozef Stefan Institute, F5, Ljubljana, Slovenia; 2 Universitat Politècnica 

de Catalunya, Departament de Física i Enginyeria Nuclear, Barcelona, 

Spain; 3 Kent State University, Liquid Crystal Institute, Kent, United States 

The addition of non-mesomorphic solutes to a nematic liquid crystal 

significantly reduces its nematic-to-isotropic transition temperature. 

Such a system is studied by using a photoisomerizable liquid crystal 

diheptylazobenzene (7AB) where the concentration of non-mesomorphic 

molecules can be controlled dynamically by changing the intensity of UV 

light. Under illumination the molecules are converted from straight trans 

to bent cis form which behaves as impurity. The resulting binary mixture 

shows a phase diagram with nematic (N), isotropic (I) and coexistence 

(N+I) region. 

Contrary to the previous measurements where the temperature 

dependence at fixed concentrations was studied, the in-situ UV 

illumination setup allowed us to measure the concentration dependence 

at fixed temperatures. The system was illuminated for a certain time 

after which the measurements of the order parameter started while the 

ABSTRACTS POSTER


system thermally relaxed back to the trans state. 

The order parameter of the nematic part of the mixture was measured 

by 2 H NMR. 7AB molecules were deuterium-labeled on α position. 

The measured frequency splitting directly corresponds to nematic 

order parameter Δν=3 ν q S (3cos 2 θ-1)/4, where ν q is the averaged 

quadrupole coupling constant, S the order parameter and θ the angle 

between nematic director and the direction of the magnetic field. 

In the isotropic phase of 7AB confined to cylindrical Anopore 

membranes, UV-illumination controlled surface induced order was 

observed. The concentration of cis-7AB can be related to the nematic 

ordering at the surface. Finally, the phase diagram of the mixture was 

determined by integrating the areas of nematic and isotropic peaks. 

The measurements were modeled phenomenologicaly with modified 

Landau-de Gennes theory. The inclusion of the concentration terms and 

spatially variable order parameter in the free energy predicts correctly 

both the bulk nematic and surface induced nematic ordering. 

Sn46 

Compensation of field distortion in high field superconducting 

magnets for solid-state NMR 

Shinagawa, Hideyuki 1 ; Ohki, Shinobu 1 ; Fujito, Teruaki 2 ; Shimizu, Tadashi 1 

1 National Institute for Materials Science, Advanced Nano 

Characterization Center, Tsukuba, Japan; 2 Probe Laboratory Inc., 

Tsukuba, Japan 

Field distortion that is induced by shimming through a coupling of the 

shim-coil and the magnet was investigated for a high magnetic field 

(1H - 930 MHz) superconducting magnet for high-resolution solid-state 

nuclear magnetic resonance (NMR). We found the field distortion or 

field drift following from the shimming, in which the shim setting was 

largely changed, may not be negligible for some experiments to obtain 

high resolution spectra using the high field superconducting magnet. 

The distortion have a relatively large time constant up to one week. 

Such a large change in the shim setting would occasionally happen 

after changing the probe. A method to computationally compensate 

such distortion in the magnetic field with an active shimming will 

be demonstrated. The external filed lock technique, we previously 

developed, in which a liquid NMR signal source is placed for field 

detection beside the solid sample, is very powerful to finely stabilize 

the field distortion but efficient only for the Z0 component. On the other 

hand, the computational method is applicable to coarsely compensate 

the distortion along all components (Z0, Z1, X1...) in the shim-coil. 

Sn47 

Ionization versus Hydrogen Bonding – How to interpret changes 

of 31 P Chemical Shift Tensor (CST) parameters for bioorganic 

compounds containing phosphoryl group 

Gajda, Jaroslaw 1 ; Olejniczak, Sebastian 1 ; Bryndal, Iwona 2 ; Potrzebowski, 

Marek 1 

1 The Centre of Molecular and Macromolecular Studies, Polish Academy 

of Sciences, Lodz, Poland; 2 University of Wroclaw, Faculty of Chemistry, 

Wroclaw, Poland 

The phosphoryl group is one of the most important functional molecules 

of biological systems, being involved in numerous processes of living 

organisms. At physiological pH, phosphate residues usually carry a 

-2 charge. The change of local electrostatic potential often induces 

conformational changes of proteins, influencing their functions, or 

modulates protein-protein interactions. Understanding the nature of 

these two factors—protonation and hydrogen bonding—is crucial for 

explaining the phosphate group’s biological function. In our project 

solid state NMR spectroscopy and gauge including atomic orbital 

(GIAO) theoretical calculations were employed to establish structural 

110 

restraints (ionization, hydrogen bonding, spatial arrangement) for 

O-phosphorylated L-threonine derivatives in different ionization states 

and hydrogen bonding strengths. Both the experimental and the GIAO 

approach show that 31 P δii chemical shift tensor parameters are very 

sensitive to ionization state. The negative values found for the skew 

κ are typical for –2 phosphates. The span Ω reflects the strength 

of hydrogen bonding. For species in –1 ionization state, engaged in 

very strong hydrogen bonds, Ω is smaller than for a phosphate group 

involved in weak hydrogen bonding. For phosphates in –2 ionization 

state, Ω is significantly smaller compared to –1 species, although the 

κ for -1 samples never reaches negative values. For –1 phosphate 

residues, in the case when 1H one pulse and/or combined rotation and 

multiple pulse spectroscopy (CRAMPS) sequences fail and assignment 

of proton chemical shift is ambiguous, a combination of 1H-13C and 

1H-31P 2D heteronuclear correlation (HETCOR) correlations is found to 

be an excellent tool for the elucidation of 1H isotropic chemical shifts. 

In addition, a 2D strategy using 1H-1H double quantum filter (DQF) 

correlations is useful for analyzing the topology of hydrogen bonding. 

In the case of a multi-center phosphorus domain, 2D 31P-31P PDSD 

experiment gives information about the spatial arrangement of the 

phosphate residues. 

Sn48 

NMR study of xenon and d-chloroform solutes in a thermotropic 

biaxial nematic liquid crystal 

Jokisaari, Jukka 1 ; Tallavaara, Pekka 1 ; Mehl, Georg 2 

1 University of Oulu, Department of Physical Sciences, Oulu, Finland; 

2 The University of Hull, Department of Chemistry, Hull, United Kingdom 

The molecules which form mesogenic phases are not usually 

cylindrically symmetric. Consequently, one could expect that apart 

from a uniaxial nematic phase, there should be a biaxial nematic 

phase. In fact, it was predicted already over 30 years ago that such 

a phase indeed should exist but only about 5 years ago first reports 

on thermotropic biaxial nematic liquid crystals were reported.1,2 The 

characterization of the biaxial phase was performed using a partially 

deuterated liquid-crystalline probe and detecting 2 H NMR spectra of a 

rotating sample, the rotation axis being perpendicular to the external 

magnetic field. 

In the present case, we propose and test an alternative NMR method. 

That is NMR spectroscopy of dissolved xenon-129 gas and deuterated 

chloroform. The sample used is TM35, organosiloxane tetrapode, which 

is supposed to display a biaxial nematic phase with the range from 

243 to 310 K. Static 129 Xe and 2 H NMR measurements of xenon and 

d-chloroform, respectively, clearly reveal different phases of the liquid 

crystal. 

[1] L. A. Madsen, T. J. Dingemans, M. Nakata, and E. T. Samulski, Phys. 

Rev. Lett. 92, 145505 (2004). 

[2] J. L. Figueirinhas, C. Cruz, D. Filip, G. Feio, A. C. Ribeiro, Y. Frère, T. 

Meyer, and G. H. Mehl, Phys. Rev. Lett. 94, 107802 (2005). 

[3] C. Cruz, J. L. Figueirinhas, D. Filip, G. Feio, A. C. Ribeiro, Y. Frère, T. 

Meyer, and G. H. Mehl, Phys. Rev. E 78, 51702 (2008). 

Sn49 

On coherences and pseudocoherences observed through the 

Carr-Purcell-Meiboom-Gill sequences 

Franzoni, María Belén; Levstein, Patricia R. 

Facultad de Matemática, Astronomía y Física. Universidad Nacional de 

Córdoba., Córdoba, Argentina 

Lately, quantum information has become a very interesting and growing 

field. Various physical systems and different techniques were proposed 


as potential hardware for quantum computing, and it is crucial the 

knowledge and correct understanding of the system decoherence 

processes. 

Nuclear Magnetic Resonance studies of coherence in C , 60 29Si and Y O , 2 3 

as good candidates for quantum information, showed very unusual 

behaviors. Commonly used pulse sequences for a spin-spin decay 

time (T ) measurement are the Hahn Echo, and multipulse sequences 

2 

like CPMG (with different pulse phase alternations). From analytic 

calculations it is well known that these sequences, when applied to 

solids, should give the same result. 

HE The Hahn echo experiment performed in C yielded a T ≈ 15 ms. 

60 2 

However unexpected long tails (T ≈2 s) appeared when the CPMG1 

2 

({Y,Y}) or the CP2 ({X,-X}) sequences were applied. Other unusual 

observations were oscillations in the magnetization decay for CPMG2 

({Y,-Y}) and a magnetization decay faster than 15 ms for CP1 ({X,X}). 

After a careful experimental and analytical study we were able to 

understand these phenomena. The unexpected behaviors arise on 

highly inhomogeneous lines compared with the dipolar couplings 

leading to slow spin diffusion. The inhomogeneity produces different 

tilting angles in different sites of the sample that the flip-flop interaction 

is unable to average out. As a consequence, there is an unwanted 

magnetization stored in the direction of the external magnetic field (z 

polarization). This polarization is brought back to the plane and observed 

either as a long tail or as oscillations. So, the long tails should not be 

interpreted as long coherence times. 

In this work, we present a complete experimental and numerical study 

of the magnetization behavior as a function of the interpulse separation 

for the different sequences. We observed that for longer interpulses the 

HE unusual behaviors tend to disappear leading to T . Our results are 

2 

important because the interpulse time can be directly associated to the 

effective dipolar interaction capable to average out the inhomogeneities. 

Sn50 

Characterizing polymorphism by Solid-State NMR, X-ray powder 

diffraction and factor analysis 

Urbanova, Martina 1 ; Brus, Jiri 1 ; Sedenkova, Ivana 1 ; Brusova, Hana 2 ; 

Kobera, Libor 1 

1 Institute of Macromolecular Chemistry AS CR, Prague, Czech Republic; 

2 Zentiva Group, a.s., Prague, Czech Republic 

The ability of the particular chemical substance to exist in several 

different crystal forms is called polymorphism. Polymorphs (crystal 

forms or crystal phases) are thus different crystal structures of the 

same compound, and are detected by various analytical techniques. 

They have different physical properties, chemical stability and reactivity, 

and consequently, also different pharmacokinetic profiles. The series 

of various polymorphs of atorvastatin was prepared and subsequently 

characterized by 13C CP/MAS NMR and 19F MAS NMR spectroscopy 

and x-ray powder diffraction. 19F MAS NMR measurement enables to 

obtain the spectra of the sample in short time. However, the differences 

in 19F MAS NMR spectra are very slight, difficult to recognize by simple 

analyze. That is why a factor analysis was applied to the sets of 19F and 

13C spectra and x-ray diffractograms of atorvastatin. The aim of the 

factor-analysis application is to find the similar features in the behavior 

of different data series. The results of the factor analysis were used 

to compare the data series and determined their possible correlation. 

In consequence, the atom groups influenced by each other can be 

eventually distinguished. 

Acknowledgements: The authors wish to thank the Ministry of 

Education, Youth and Sports for financial support (grant No. 2B08021). 


Sn51 

Formation of SAPO and MnSAPO thin films and xerogels studied 

by solid-state NMR spectroscopy 

Rangus, Mojca; Mali, Gregor; Cecowski, Saša; Novak Tušar, Nataša; 

Kaučič, Venčeslav 

National Institute of Chemistry, Ljubljana, Slovenia 

Transition-metal (Me) functionalized mesoporous aluminophosphate 

(AlPO) and silicoaluminophosphate materials (SAPO) have been widely 

investigated as heterogeneous catalysts due to their hydrothermal 

stability and high catalytic activity [1]. MeAlPO and MeSAPO as powders 

have attracted attention as acid, base and redox heterogeneous 

catalysts. As thin films, these materials also have potential in optics 

and electronics. Recently, the detailed preparation and structural 

characterisation of thermally stable mesoporous cubic AlPO as a thin 

film was reported for the first time [2]. 

We prepared mesoporous aluminophosphate thin films and xerogels 

with ordered cubic pore arrangement, functionalized by silicon (SAPO) 

or silicon and manganese (MnSAPO). These samples were than studied 

by solid-state NMR spectroscopy to determine the local framework 

structure. 

27 Al, 31 P and 29 Si MAS NMR spectra revealed changes in the structure 

after the samples were calcined. In both, SAPO and MnSAPO thin 

films, the fraction of tetra-coordinated aluminium increases after the 

calcination, while the peak belonging to hexa-coordinated alumina 

diminishes. This change is even more pronounced in xerogel samples. 

27 Al and 31 P NMR spectra reveal that mesoporous thin films contain 

more defect sites and are therefore less condensed than frameworks of 

xerogel samples. 

Because of the very small amount of silicon in the framework and low 

sensitivity of 29 Si nuclei, we were not able to detect 29 Si MAS NMR 

spectra of the SAPO thin films. In MnSAPO samples NMR-active nuclei 

had much shorter relaxation times and we were therefore able to 

accumulate more scans while recording the spectra. 29 Si NMR spectra 

in MnSAPO thin films, similarly as 27 Al and 31 P spectra indicated that 

the mesoporous framework was less condensed than the framework in 

xerogels, and that in all the samples the Si is incorporated in the walls 

of mesoporous material in form of silicon rich domains. 

[1] A. Corma, H. Garcia, Chem. Rev. 102 (2002) 3837. 

[2] M. Mazaj et al., Langmuir, 24 (2008) 6220. 

Sn52 

Implementation of double resonance magic angle coil spinning 

experiments 

Inukai, Munehiro; Takeda, Kazuyuki 

Kyoto University, Division of Chemistry, Graduate School of Science, 

Kyoto, Japan 

Magic angle coil spinning (MACS), put forth by Sakellariou et al., 

has opened a way to study molecular structure and dynamics in tiny 

quantities of polycrystalline/amorphous materials with increased 

sensitivity and high resolution. As reported, wireless, inductive coupling 

of a primary coil in the conventional MAS probe with a tiny rf resonator 

embedded in a rotor, and its spinning together with the sample of 

interest, not only lead to a higher filling factor, but also beautifully 

solve the problem intrinsic to the stationary microcoil of static-field 

distortion due to the bulk susceptibility effect of the coil material itself. 

Also importantly, MACS, unlike the other strategies for microcoil-MAS 

reported so far, does not require any hardware modification to the 

existing MAS probe/module, attracting NMR users (chemists and 

biologists) as well as NMR developers. Since MACS has currently been 

demonstrated in single resonance experiments, a question arises; 

is it also applicable to double resonance experiments, which would 

ABSTRACTS POSTER


undoubtedly be beneficial? 

In this work, we demonstrate 1H-13C double-resonance MACS 

experiments. In order to handle two separate frequencies, we fabricated 

a doubley-tuned MACS resonator (resonant frequency: 75 MHz and 300 

MHz) composed of two microcoils (i.d. : 0.5 mm) and two capacitors to 

be spun together with the rotor. The double-resonance MACS scheme 

allows us to implement powerful double-resonance solid-state NMR 

methodologies including cross polarization, proton decoupling, and 2D 

correlation spectroscopy etc., while still enjoying the merits of MACS, 

such as intense rf irradiation, high-sensitivity, eliminated magnetic 

susceptibility-induced field distortion, and an easy-to-use approach with 

the conventional and widespread hardware. 

Sn53 

HRMAS NMR for the investigation of grafted organotin catalysts 

at the solid-liquid interface 

Pinoie, Vanja; Biesemans, Monique; Willem, Rudolph 

Vrije Universiteit Brussel, High Resolution NMR Centre, Brussels, 

Belgium 

Tin-based Lewis acids such as mono- or dialkyltin compounds are 

efficient transesterification catalysts. Nevertheless, their routine use in 

an industrial context has been limited so far by their intrinsic toxicity 

and by the generally tedious quantitative removal of tin-containing 

reagents or by-products from the final reaction product. A scientifically 

rewarding strategy to overcome the latter concerns involves grafting of 

the organotin reagent onto an insoluble support. 

The synthesis and catalysis of a functionally pure monoalkyltin 

trichloride grafted onto cross-linked polystyrene are monitored by 

High-Resolution Magic Angle Spinning (HRMAS) NMR combined with 

119 Sn based 1D and 2D NMR techniques. [1,2] This characterization tool 

provides high-resolution NMR spectra of solid-supported functional 

groups in situ at the solid-liquid interface, the support being immersed 

in a solvent. Diffusion-filtered 1 H HRMAS NMR of the reaction mixture 

allows investigating the catalyst in situ during the transesterification 

by discriminating between the catalytic grafts and the translationally 

mobile reaction mixture components, and provides, together with 119 Sn 

HRMAS NMR, a better insight into the catalysis mechanism. Quantitative 

determination of the tin loading is performed using a calibrated 

electronically produced signal as an internal reference, i.e. ERETIC, 

applied to 119 Sn HRMAS NMR. [3] 

In a study in which the spacer is tuned toward higher polarities, clearly 

identifiable tin coupling patterns in the 2D 1 H- 13 C HSQC spectra provide 

valuable information with regard to the extent of tin coordination 

in these materials and point toward the existence of intra- and/or 

intermolecular O→Sn interactions due to the presence of donor moieties 

in the linker. 

Switching to inorganic catalyst carriers, 29 Si solid-state NMR and 1D 

and 2D 1 H, 13 C and 119 Sn HRMAS NMR evidence their full potential 

as complementary analytical tools in the thorough characterization of 

silica-supported organotin grafts and their precursors. 

[1] J. C. Martins et al., Chem. Eur. J. 2002, 8, 3431. 

[2] V. Pinoie et al., Organometallics 2007, 26, 6718. 

[3] V. Pinoie et al., Organometallics 2008, 27, 3633. 

112 

Solution NMR 

So10 

Investigation of alcoholic acidic aluminophosphate solution 

using 27Al and 31P NMR spectroscopy 

Samadi-Maybodi, Abdolraouf 1 ; Hassani Nejad, Sayed Karim 2 

1 Faculty of Chemistry, University of Mazandaran, Chemistry, Babolsar, 

Islamic Republic of Iran; 2 University of Mazandaran, Chemistry, Babolsar, 

Islamic Republic of Iran 

Phosphorus-31 and aluminum-27 nuclear magnetic resonance 

techniques were used to characterize the distribution of soluble 

aluminophosphate species in aqueous solutions of (2-hydroxyethyl) 

trimethylammonium chloride (2-HETMACl), phosphoric acid, and 

aluminum sulfate [1]. Soluble aluminophosphate cations form reactions 

of [A1(H2O)6]3+ with phosphate ligands (i.e., H3PO4, H2PO4¨C, and 

acid dimers H6P2O8 and H5P2O7¨). Aluminophosphate species, such 

as [Al(H2O)4,(OH)(H2PO4)]+, have been suggested as the nutrients 

for the growth of AlPOs molecular sieves [2]. It was proved the 

presence of [Al(H2O)5(H3PO4)]3+, [Al(H2O)5(H3PO4)n]m+ (n≥2, m 

is undetermined), [Al(H2O)5(H2PO4)]2+ and [A1(H2O)4(H2PO4)2]+ 

cations under acidic conditions [3]. 

In this work, we investigated the effect of poly ethylene glycol (pEG) and 

methanol in formation of these cations. New information was obtained 

from this study indicating aluminophosphate species in the mixed 

solvent of H2O/Methanol or H2O/pEG. Results indicated that 31P and 

27Al signals enhanced with above mixed solvent and also in the H2O/ 

Methanol solvent, all signals more enhanced than theirs in the H2O/ 

pEG solvent. These results also can help to better understanding of the 

synthesis of the AlPO4 zeolites in non-aqueous media. 

References 

[1] A. Samadi¨CMaybodia,*, S.K. Hassani Nejad¨CDarzia and H.R. 

Bijanzadeh, Spectrochim Acta A, 72 (2009) 382¨C389 

[2] X. Ren, S. Komarneni, D.M. Roy, Zeolites, 11 (1991) 142¨C148. 

[3] R.F. Mortlock, A.T. Bell, C.J.R. Radke, J. Phys. Chem. 97 (1993) 

767¨C774. 

So11 

Investigation of solvation properties of organic molecules in 

aqueous solutions by nmr - relaxation method 

Donets, Alexey; Chizhik, Vladimir 

Saint-Petersburg State University, Physical department, Saint- 

Petersburg, Russian Federation 

The ion-protein interaction has long been recognized as a major 

determinant of chain folding, conformational stability, internal dynamics 

and binding specificity of globular proteins. 

The method developed in the Department of Quantum Magnetic 

Phenomena (Saint-Petersburg State University) makes it possible to 

determine important parameters of the microstructure of solutions using 

measurements of magnetic relaxation rates of solvent and solute nuclei 

as functions of concentration and temperature. It was experimentally 

shown, that the majority of ions maintain their coordination number in 

variations of concentration and the temperature in a wide range. But 

the study of electrolyte solutions containing anions (Br - and Cl - ) has 

shown that the coordination number of these anions may vary due to 

temperature variations [1]. The changes of the anions hydration shell in 

the interval 30

solvated chlorine ion in the aqueous salt solution containing organic 

molecules. The method is based on the analysis of temperature 

dependences of magnetic relaxation rates of the 35 Cl nuclei in the 

simple electrolyte solution and with proteins. The method is simple and 

does not demand use of complex pulse sequences. 

Acknowledgements 

This work was supported by the Russian Foundation for Basic Research, 

project No 07-03-00735. 

References 

[1] A.V. Donets, V.I. Chizhik. Rus. J.Phys.Chem. Vol. 79, 1 6, 2005, p. 

1032-1036 

So12 

NMR as a tool to study free and adsorbed phosphorus species 

in boreal ecosystems 

Vincent, Andrea 1 ; Sleucher, Jürgen 2 ; Gröbner, Gerhard 3 ; Persson, Per 3 ; 

Jansson, Mats 1 ; Giesler, Reiner 1 

1 Umeå University, Ecology and Environmental Science, Umeå, Sweden; 

2 Umeå University, Medical Biochemistry and Biophysics, Umeå, Sweden; 

3 Umeå University, Chemistry, Umeå, Sweden 

Phosphorus (P) is globally a key element affecting plant productivity in 

agricultural and natural ecosystems. Organic P is the dominant P form 

and is associated with the soil organic matter and the dissolved organic 

matter in soil, stream and lake water. Knowledge on the speciation 

and reactivity of organic P is thus key for both terrestrial and aquatic 

science, and even to future policy making, as it is necessary to predict 

the impacts of P inputs to the Baltic Sea, and of global anthropogenic 

change on the P cycle and hence plant productivity. Despite its 

importance, our advances in the speciation of organic P during the last 

20 years are scant. 

In this contribution we compare NMR methods to analyze the speciation 

of organic P in soils and freshwater systems. We also give preliminary 

NMR results on P speciation in northern forests, and on molecular 

details of how different organic P species relate to vegetation, climate 

and microbial bioavailability. We show that P limitation in certain forests 

is likely to arise due to shifts in P speciation relative to non-P limited 

sites, rather than to a decrease in the total P content of soils. Liquid 

state 1D 31P NMR is the most commonly used NMR experiment for the 

analysis of organic P. While this experiment resolves several classes 

of P compounds, it suffers from important drawbacks: Firstly, organic 

P must be extracted, which means that results depend strongly on the 

extractant used. Secondly, the alkaline conditions necessary to optimize 

resolution result in the hydrolysis of certain compounds during analysis. 

Thirdly, the identification of specific P monoesters and P diesters is 

difficult as many signals are ‘buried’ under a single peak. 

We are addressing these problems with: 

- Alternative sample preparation methods 

- 2D NMR, which increases sensitivity and resolves P species by 31P 

and 1H chemical shifts 

- The use of solid-state NMR, which does not require an extraction step, 

and enables the building of P mass balances 

In summary, advanced NMR methods increase the information on 

P species in the environment and their turnover, helping to predict 

ecosystem response to global anthropogenic change. 


So13 

Biophysical characterization of neuronal calcium sensor 1, a 

comparative study in myristoylated and non-myristoylated form 

Kousik, Chandra 1 ; Sharma, Yogendra 2 ; Chary, Kandala V 3 

1 Tata Institute of Fundamental Research, Cemical Science, Mumbai, 

India; 2 Centre for Cellular and Molecular Biology, Biological Science, 

Hyderabad, India; 3 Tata Institute of Fundamental Research, Chemical 

Science, Mumbai, India 

Neuronal calcium sensor-1 (NCS-1) is a major modulator of Ca 2+ - 

signaling with a known role in neurotransmitter release. The 15N-1H 

HSQC spectra of myristoylated and the non-myristoylated NCS1 reveals 

that the backbone structures of these two proteins are not drastically 

different. However, Ca 2+ -filling pathways are different in the two forms. 

This suggests that myristoylation influences the protein response to 

alteration in Ca 2+ concentration, that is crucial for its physiological 

functions. For understanding the specialized functions of NCS-1, 

identification of ionic specificity of individual sites is important. In this 

study, we have identified the specificity of Ca 2+ -binding and the role of 

Mg 2+ in modulating Ca 2+ -binding to NCS-1 by NMR. Our data suggests 

that Mg 2+ binds only to the EF2 and EF3 and thus classifying them 

as structural sites, while EF4 is a Ca 2+ -specific or regulatory site. In 

the presence of Mg 2+ , Ca 2+ -binding induces unusual conformational 

rearrangements in the protein, and Ca 2+ reverses the Mg 2+ , induced 

changes. In a larger physiological perspective, the reduction in the 

overall affinity of Ca 2+ in the presence of Mg 2+ would be advantageous 

to the molecule by facilitating the reversibility to its Ca 2+ -free state. 

We have also characterized of the near native energy states of NCS- 

1 in both myristoylated and non-myristoylated forms and the effect 

of myristoylation in determining the extent of ruggedness due to 

the accessing of various alternative states, which is crucial for the 

functionality of the protein. 

So14 

EASY ROESY, Reliable cross peak integration in adiabatic 

symmetrized ROESY 

Thiele, Christina 1 ; Petzold, Katja 2 ; Schleucher, Jürgen 2 

1 Technische Universität Darmstadt, Clemens Schöpf Institut, Darmstadt, 

Germany; 2 Umea University, Medical Biochemistry and Biophysics, 

Umea, Sweden 

The determination of the three dimensional structure of organic and 

biomolecular compounds by NMR spectroscopy usually relies on 3 J 

couplings and NOEs. The NOE, however, changes sign at ωτ c = 1.12, 

such that little or no NOE is observed for medium sized compounds 

(MW ca. 1000 Da). This problem can be solved by using ROESY, which 

yields negative cross-peaks for all values of ωτ c . 

In the interpretation of ROESY, however, several experimental problems 

- namely COSY-type artefacts, Hartmann-Hahn matching and offset 

dependence - have to be avoided. These lead to serious complications, 

when distances or exchange rates are to be extracted from ROESY 

spectra. 

Several ROESY pulse sequences have been proposed for removing 

these artefacts[1], but they are either cumbersome to set up, reduce 

sensitivity too much, or show pronounced off set dependence. 

We show that by bracketing two off-resonance spin-lock pulses 

between adiabatic pulses, we obtain high-quality, high sensitivity 

spectra with negligible offset-dependence, without need for samplespecific 

set up. The resulting spectra can be reliably quantified and 

TOCSY transfer is reduced almost to its theoretical limit. The superior 

performance of the EASY-ROESY [2] (Efficient Adiabatic SYmmetrized 

ROESY) is demonstrated by comparing ROESY and NOESY-derived 

distances for a small protein (BPTI) and by measuring exchange rates in 

an organometallic (Grubbs II) complex. 

ABSTRACTS POSTER


References: 

[1] T.-L. Hwang, A.J. Shaka, J. Am. Chem. Soc. 1992, 114, 3157- 

3159; J. Schleucher, J. Quant, S. Glaser, C. Griesinger, J. Magn. Reson. 

A, 1995, 112, 144-151; H. Desvaux, P. Berthault, N. Birlirakis, M. 

Goldman, M. Piotto, J. Magn. Reson. A 1995, 113, 47-52. 

[2] C. M. Thiele, K. Petzold, J. Schleucher, Chem. Eur. J. 2009, 15, 585- 

588. 

So15 

Structural insight into Myb-like domain of telomeric repeat 

binding protein derived from zebra fish 

Yun, Ji Hye; Lee, Inhwan; Kwak, Sooyoung; Kim, Sangyun; Ko, 

Sunggeon; Lee, Weontae 

Yonsei University, Biochemistry, Seoul, Republic of Korea 

Telomeres are repetitive DNA sequences located at the termini of 

linear chromosomes, which protects the end of the chromosome from 

destruction. They consist of tandem repeats of TTAGGG in vertebrates 

or TTTAGGG in higher plants. The myb-like domain which involved in 

telomere binding protein recognizes specific telomeric DNA repeat 

sequence and makes DNA complexes which may modulate the length 

of telomere. The specificity of telomere DNA binding is determined by 

the myb-like domain which is well conserved in many species. Based 

on sequence homology analysis, we identified telomere DNA binding 

domain (522-570) at C-terminus in telomeric repeat binding protein 

of zebra fish (zTerfa). It is classified as the myb-like domain compared 

with other myb-like domains, but showed low sequence similarity with 

hTRF1. In order to determine the solution structure of myb-like domain 

of zTerfa, we have cloned and expressed the DNA binding domain 

in E.coli. The conventional backbone and NOE experiments were 

performed in Bruker DRX 900MHz with Cryoprobe. NOE and TALOS 

were used for distance constraints and angle constrains, respectively. 

The solution structure was calculated using CYANA 2.2.5. version. The 

structure contains three alpha helices. Electrostatic potential surface 

shows helix 3 has positive charged amino acids, which might take 

part in DNA-protein interaction. We have performed isothermal titration 

calorymetry (ITC) and NMR titration experiments to investigate the DNA 

binding mechanism of zTerfa. The results indicate that the DNA binding 

domain of zTerfa interacts not only with human telomere repeats but 

also with plant telomere repeats as 2:1 molar ratio and have different 

DNA binding modules with each human and plant telomere repeats. 

So16 

Studies of macromolecular interactions of glucose-dependent 

insulinotropic polypeptide and its receptor based on NMR, 

molecular modelling and docking approaches 

Venneti, Kalyana 1 ; O’Harte, Finbarr 2 ; Hewage, Chandralal 1 

1 University College Dublin, School of Biomolecular and Biomedical 

Science, Dublin, Ireland; 2 University of Ulster, School of Biomedical 

Sciences, Coleraine, Northern Ireland, United Kingdom 

Glucose-dependent insulinotropic polypeptide (GIP) is a gastrointestinal 

hormone that enhances glucose stimulated insulin secretion by 

interacting with a hetero-trimeric G-protein coupled receptor located 

in pancreatic β-cell. Due to its glucose lowering and insulinotropic 

properties, GIP is considered as a potential target for treating type II 

diabetes. Being involved in the drug development research for type II 

diabetes, we were interested to study the importance of structure of 

GIP and its analogues in determining their biological activity. In our 

laboratory, we identified the solution structures of GIP in membrane 

mimicking (micellular and bicellular) medium using NMR, computational 

modelling techniques and also determined the receptor-hormone 

interactions between the GIP ligand and N-terminal extra-cellular 

114 

domain (N-ECD) of the GIP receptor using macro-molecular docking 

approaches. The results of this study indicate that the docking 

conformation of GIP is stabilised by the presence of hydrophobic and 

intermolecular hydrogen bonding interactions. The ligand-receptor 

complex model highlights the possible interactions between the 

residues Glu19, Gln20, Lys30 and Lys33 of the GIP-ligand with the 

residues Gln30, Asn120, His115 and Leu111 of the N-terminal domain 

of the GIP receptor respectively. This information is useful and vital for 

understanding the biologically relevant receptor-bound structure of GIP 

that facilitates the design of new peptide or non-peptide based drugs 

for treating type 2 diabetes and other related disorders. 

So17 

Multiple-Encoded HSQC-TOCSY experiment 

Gil, Sergi; Pérez, Miriam; Nolis, Pau; Parella, Teodor 

Universitat Autònoma de Barcelona, Servei de Ressonància Magnètica 

Nuclear, Bellaterra, Barcelona, Spain 

Heteronuclear-edited correlation experiments of the type 2D 1H–X HSQC–TOCSY have been developed into powerful NMR tools for 

structural studies of molecules in which the analysis of conventional 

TOCSY spectra can fail due to undesired overlapping effects. 1 

A new strategy to obtain multiple-encoded HSQC-TOCSY spectra is 

here proposed. We evaluate the possibility to simultaneously include 

different types of signal editing into the original HSQC-TOCSY pulse 

sequence (IPAP editing, direct vs relayed responses or multiplicity 

editing). Equivalent and complementary NMR data are acquired and 

processed using Hadamard-type protocols. 2 The information usually 

coded in a single HSQC-TOCSY spectrum can thus be separately 

obtained from eight edited NMR spectra allowing better performance 

when resonance overlapping could prevent good data interpretation. In 

this way, information about carbon multiplicity, differentiation between 

direct HSQC vs relayed TOCSY cross peaks, and measurement of the 

sign and the magnitude of both large, direct and small, long-range 

proton-carbon coupling constants can be simultaneously extracted from 

this single NMR experiment. 

Selective and band-selective 2D ME-HSQC-TOCSY experiments, as 

well as 1D and 3D versions, will proposed as useful tools for structural 

elucidation of small molecules. The method will be also applied to 

measure the sign and the magnitude of small, long-range protoncarbon 

coupling constants. The application of the concepts in timeshared 

13C/ 15N-HSQC-TOCSY 3 experiments will also permit the study 

on nitrogen-containing compounds at natural abundance. Examples 

showing the advantage to avoid accidental signal overlapping due 

to the satellite line of the direct responses or carbon chemical shift 

degeneracy are provided. 

1 P. Nolis, T. Parella. J. Magn. Reson. 2005; 176 : 15-26 

2 E..Kupce, T. Nishida, R.Freeman. Prog. NMR Spectrosc. 2003; 42: 95 

K.Krishnamurthy. J.Magn.Reson. 2001; 153: 144-150 

3 P. Nolis, M. Pérez, T. Parella. Magn. Reson. Chem. 2006; 44: 1031- 

1036 

M. Sattler, M. Maurer, J. Schleucher,C. Griesinger. J. Biomol.NMR 1995; 

5: 97-102 


So18 

H(C)H experiment: extending Long- range 1 H- 1 H connectivities 

via an intermediate heterospin 

Gil, Sergi; Pérez, Miriam; Nolis, Pau; Parella, Teodor 

Universitat Autònoma de Barcelona, Servei de Ressonància Magnètica 

Nuclear, E-08193-Bellaterra, Barcelona, Spain 

The structural characterization of small and medium-sized molecules 

by high-resolution NMR spectroscopy is routinely made by analysis of 

through-bond and through-space interactions extracted from a variety 

of J-based and nuclear overhauser effect (NOE)-based multidimensional 

NMR experiments, respectively. The most characteristic two- and threebond 

homo- and heteronuclear connectivities are mainly determined 

from COSY 1 and HMBC 2 spectra and only in exceptional cases or using 

modified sequences, longer interactions can also be ascertained. 

Instead of direct J-coupling between two protons, which in the case 

of many bonds separation would be too small, we propose a new type 

of proton–proton correlation experiment to establish multiple-bond 

connectivities via an intermediate 13 C heteronucleus, as an alternative 

way to avoid carbon overlapping in the commonly used HMBC 

experiment. The magnetization flow pathway is based on a consecutive, 

dual-step J(CH)–transfer mechanism and it allows one to trace out 1 H– 

1 H connectivities between protons belonging to different spin systems. 

In favourable and specific spin topologies, such as those containing 

ϖ systems showing considerable 4 J(CH) or 5 J(CH) coupling values, it 

should be expected that reliable information could also be detected 

between protons separated up to seven or eight bonds. This novel H(C) 

H experimental scheme will be particularly useful in cases when carbon 

resonances overlap, providing connectivity information that could not 

be obtained in a HMBC experiment 3 . Selective 1D and general 2D 

versions will be presented and illustrated on small molecules at natural 

abundance. 

1U. Piantini, OW. Sørensen, RR. Ernst. J.Am.Chem.Soc. 1982; 104: 

6800. 

2A. Bax, MF. Summers. J. Am. Chem. Soc. 1986; 108: 2093. 

3 T. Parella, J.F. Espinosa. Magn. Reson. Chem. 2008; 44: 464-470. 

So19 

The solution structure of a novel protein containing a new 

heterometallic molybdenum-copper cluster 

Pauleta, Sofia R. 1 ; Carepo, MS 1 ; Turano, P. 2 ; Bertini, I 2 ; Moura, JJG 1 ; 

Moura, I 1 

1 REQUIMTE, CQFB, Dep. Química, FCT-UNL, Caparica, Portugal; 2 CERM, 

Via Sacconi 6, Sesto Fiorentino, Italy 

The ORange Protein (ORP) from Desulfovibrio gigas is an orange 

coloured 11.8 kDa protein that contains a mixed-metal sulphide cluster, 

of the type [S2MoS2CuS2MoS2]3-, non-covalently bound to the 

polypeptide chain [1,2]. A blast search revealed that this protein has 

sequence homology of around 30 to 50 % with conserved proteins from 

eubacteria and hyperthermophilic archaea with unknown function. They 

all contain a conserved domain common to the nitrogenase accessory 

factor (NifB C-terminal domain, NifX and NafY). The ORP was produced 

for NMR studies by heterologous expression in E. coli as the apo-form 

[3]. The holo form was reconstituted by the in situ synthesis of the 

metal cluster upon the addition of copper sulphate and thiomolybdate 

or thiotungstate. The polypeptide chain of ORP is composed of 117 

residues of which 5 are prolines. 13C detection experiments enabled 

the extension and confirmation of the sequential assignment for both 

apo and reconstituted-forms of ORP [3]. The over-all solution structure 

of the reconstituted ORP has a α/β motif, characteristic of the members 

of the ribonuclease H family. The mapping of the chemical shift 

differences between apo and reconstituted form was used to elucidate 

which region of the polypeptide chain is involved in the binding of 

the metal cluster, which coincide with mobile regions in the apo-ORP 

identified in 15N relaxation experiments. 

These results give insights into the metal binding mode of chaperons 

involved in the synthesis of the nitrogenase metal cofactor. 

1- G.N. George, et al. JACS, 122, 8321-23 (2000). 

2- S.A. Bursakov, et al. JIB. 98, 833-837 (2004). 3- S.R. Pauleta, et al. 

Biomol. NMR Assignm., 1, 81-83 (2007). 

This work was supported by the project POCI/QUI/55350/2004 

(Fundação para a Ciência e Tecnologia) and by the EU-NMR Research 

Infrastructure (Contract n° 026145). 


So20 

NMR sampling of the near native states in the native–state 

ensemble of dynein light chain dimer 

Chakraborty, Swagata; Krishnamohan, Poluri Maruthi; Hosur, 

Ramakrishna.V 

Tata Institute of Fundamental Research, Chemical sciences, Mumbai, 

India 

Characterizing the near-native states in the folding energy landscape of 

a protein is an exciting topic in structural biology as this manifests the 

adaptability of the protein structure to external perturbations thereby 

throwing light into its functional relevance. Here, we have investigated 

the near native states, created by sub-denaturing concentrations of 

urea in the dynein light chain protein (DLC8), the smallest subunit of 

the dynein motor complex which acts as a cargo adaptor. Equilibrium 

unfolding experiments at pH 7 using both circular dichroism and 

fluorescence spectroscopy suggested a simple two state unfolding 

of DLC8 dimer with a transition midpoint at ~ 8.5 M. The unfolding 

transition is sharp with a flat baseline up to 7M Urea. The 1H-15N 

HSQC spectrum displayed additional peaks (corresponding to monomer 

and unfolded species) at 8.0 M urea suggesting presence of alternative 

conformations in slow exchange with the major dimeric species. The 

15N-1H chemical shift perturbations as a function of urea concentration 

in the range 1-6M, revealed discernible structural change in the 

secondary structural elements in the dimer. The 13C (Cá, Câ, and CO) 

chemical shifts provided insight into the change in the torsion angles 

leading to the near native states which are scarcely populated in the 

sub-denaturing urea condition. Furthermore, the loss of protection in the 

secondary structural elements like á1 and á2 helix; â4 and â5 sheets 

clearly indicates progressive increase in the solvent accessibility of the 

dimeric protein at higher urea concentrations. R1, R2 and heteronuclear 

NOE were measured for the protein at 0-6M urea, to probe the changes 

in the backbone dynamics in the near native states. The above studies 

provide valuable information about the interaction potential energy 

landscape of the DLC8 dimer which is crucial for understanding its 

interaction with different targets. 

So21 

Allosteric mechanism of Hsp70 molecular chaperones 

Zhuravleva, Anastasia; Clerico, Eugenia M.; Smock, Robert G.; Gierasch, 

Lila M. 

University of Massachusetts, Amherst, Department of Biochemistry & 

Molecular Biology, Amherst, United States 

Hsp70 molecular chaperones comprise one of the major classes 

of heat-shock proteins and facilitate protein folding, disaggregation 

and translocation in all organisms. The C-terminal substrate-binding 

domain (SBD) of Hsp70s specifically recognizes and binds to extended, 

hydrophobic regions of partially unfolded protein substrates. ATP binding 

to the nucleotide-binding domain (NBD) induces substrate release, 

ABSTRACTS POSTER


allowing for further cycles of refolding. Although structural information is 

available for the isolated NBD and SBD and for ADP-bound two-domain 

constructs of Hsp70s, the conformational changes associated with their 

allosteric regulation remain largely mysterious. We performed a solution 

NMR study of the Escherichia coli Hsp70 homologue DnaK both as a 

two-domain protein and as isolated domains to deduce conformational 

changes in its allosteric cycle. Our data provide, for the first time, 

atomic detail descriptions of the low-affinity ATP-bound conformation of 

DnaK in the presence and absence of the substrate. Our NMR results 

for different constructs of the isolated NBD together with statistical 

coupling analysis (SCA) and MD simulations reveal the network of 

residues involved in the allosteric response. Taken together, our data 

we suggest the following mechanism of allostery: ATP binding induces 

conformational changes concomitant with sub-domain re-orientations in 

NBD. Interactions of the interdomain linker with the NBD are essential to 

stabilize the conformation favorable for ATP hydrolysis. However, in the 

absence of substrate, the C-terminal lid of the SBD interacts with NBD 

sub-domains IA and IB, which prevents high ATPase activity and leads 

to significant changes in the substrate-binding pocket and substantially 

increased flexibility throughout the entire SBD. When the substrate 

binds, the helical lid undocks from the NBD and the equilibrium shifts 

to the conformation favorable for ATP hydrolysis. This series of events 

offers a structural explanation for substrate-mediated enhancement 

of ATPase activity, and ATP-mediated reduction of substrate affinity, 

the central hallmarks of allostery in Hsp70s. [Supported by NIH grant 

GM027616] 

So22 

Structure and function of Mia40, a key player in mitochondrial 

oxidative folding with a novel CPC active site 

Gallo, Angelo 1 ; Banci, Lucia 1 ; Bertini, Ivano 1 ; Cefaro, Chiara 1 ; Ciofi- 

Baffoni, Simone 1 ; Martinelli, Manuele 1 ; Sideris, Dionisa 2 ; Katrakili, Nitsa 2 ; 

Tokatlidis, Kostas 2 

1 University of Florence, CERM, Florence, Italy; 2 Institute of Molecular 

Biology and Biotechnology, Foundation for Research and Technology 

Hellas, Heraklion, Greece 

Mia40 (Mitochondrial Import and Assembly) is a protein of 16 kDa of 

the intermembrane space of mitochondria (IMS) which plays a key role 

in the IMS. Mia40 is indeed part of a complex machinery involved in 

oxidadive folding of Cys-rich protein once imported in the IMS. It has 

been demostrated that Mia40 has several kind of substrates having 

or CX9C motifs, like Cox17, or CX3C motifs, like TIMs, interacting with 

them via disulphide bond. 

The solution structure of Mia40 is available. The aim of this work was 

to solve the solution structure of this protein, characterize its backbone 

dynamic and investigate the oxidative folding mechanism at the 

molecular level through NMR. The solution structure of Mia40 consists 

of a CHCH (coiled coil-helix-coiled coil-helix) domain stabilized by two 

disulfide bonds, preceded by a partially structured N-terminal tail (lid), 

containing a fully conserved CPC motif, and followed by a completely 

unstructured, flexible C-terminal tail. The two cysteins in the N-terminal 

tail (Cys 53 and Cys 55) are easily reduced at variance of the other 

two disulphide bonds. Protein-protein interaction studies of Mia40 with 

its protein partner, the copper chaperone Cox17 which is necessary 

for CcO assembly, shows that Mia40 is capable to catalyze the 

formation of the functionally active state of Cox17 where two disulphide 

bonds are formed. The CPC active site is solvent-accessible, with its 

second cysteine adjacent to a putative substrate-binding hydrophobic 

cleft created by conserved aromatic and hydrophobic residues that 

connect the lid to the core. C55 is essential in vivo and critical for 

mixed disulfide formation with the substrate. Mia40 plays a key role 

in oxidative protein folding in the mitochondrial IMS, a process not yet 

understood. This work elucidates for the first time how the oxidative 

folding mechanism operates at the molecular level in the IMS. 

116 

So23 

Structural studies of frog skin peptide XT-7 and its analogues 

using solution state NMR and molecular modelling 

Subasinghage, Anusha Priyangika 1 ; Hewage, Chandralal 1 ; Conlon, 

Michael 2 

1 University College Dublin, UCD School of Biomolecular and biomedical 

Science, Dublin 4, Ireland; 2 United ArabEmirates University, Department 

of Biochemistry, 17666 Al-Ain, United Arab Emirates 

XT-7 is a short antibacterial peptide, isolated from the skin of frog 

Xenopus tropicalis composed of 18 amino acids. It showed a broad 

spectrum of antibacterial activity against a range of reference strains of 

both Gram-negative and Gram-positive bacteria and yeast pathogens, 

but its therapeutic potential is limited by toxicity against mammalian 

cells. Substitution of Gly4 by L-Lys in XT-7 produced an analogue with 

high potency against microorganisms but low cytolytic activity in human 

erythrocytes. Therefore these peptides would be considered as the 

potential for development in to therapeutically valuable antimicrobial 

agents. 

To understand the basic structural requirements for the biological 

activity of XT-7 and its Lys4 substituted peptide, the solution structure 

was investigated by proton NMR spectroscopy and molecular modelling 

in various solvent systems. The structures of XT-7 and its analogue are 

characterized by á-helical conformation between Gly4-Val13 and Pro5- 

Gly14, respectively. 

Structural information obtained from this project could help to 

understand the structure activity relationship of XT-7 and its analogue 

and hence to facilitate development of antimicrobial drugs. 

References 

Conlon, J. M., Galadari, S., Raza, H. & Condamine, E. (2008) Design 

of potent, non-toxic antimicrobial agents based upon the naturally 

occurring frog skin peptides, ascaphin-8 and peptide XT-7. Chem Biol 

Drug Des, 72, 58-64. 

So24 

Applications of combined liquid state NMR/UV-Vis spectroscopy 

in hydrogen bond research 

Koeppe, Benjamin; Tolstoy, Peter; Limbach, Hans-Heinrich 

Free University of Berlin, Institute of Chemistry and Biochemistry, Berlin, 

Germany 

UVNMR [1] is a technique for the simultaneous measurement of 

NMR and UV-Vis absorption spectra. It combines the high content of 

structural information of NMR spectra with the superior time resolution 

of optical spectra. It may thus provide valuable information concerning 

dynamics fast on the NMR timescale, e.g. in hydrogen bonded systems. 

The method allowed us to correlate UV-Vis absorption band shifts of 

phenols in strong hydrogen bonds with the corresponding hydrogen 

bond geometries as determined by NMR. In turn, the optical spectra 

enable us to detect short lived species such as tautomeric forms of a 

hydrogen bond and to estimate the geometries as well as the population 

ratio of these tautomers. Examples are given for intermolecular OHO 

and OHN hydrogen bonds of phenols with carboxylic acids or pyridines 

in aprotic solvents. The influence of solvent polarity on hydrogen bond 

geometries in these two classes of complexes are discussed. 

UVNMR also permits to determine the binding pattern and dynamics 

of such species as the complex of one carboxylate anion with two 

phenol molecules: the carboxylate takes the central position is hydrogen 

bonded to both phenols symmetrically. 

[1] P.M. Tolstoy, B. Koeppe, et al., Angew. Chem 2009, DOI: 10.1002/ 

anie.200806181 


So25 

Automated processing of NMR data with coupled evolution 

periods for high-throughput protein analysis 

Staykova, Doroteya 1 ; Fredriksson, Jonas 1 ; Bermel, Wolfgang 2 ; Billeter, 

Martin 1 

1 University of Gothenburg, Biophysics Group, Göteborg, Sweden; 

2 Application Lab, Bruker BioSpin GmbH, Rheinstetten, Germany 

New developments of the software tool PRODECOMP 1-3 for 

decomposition of projection NMR data will be presented. Assignment 

of backbone resonances from decompositions using a routine called 

SHABBA (Shape Analysis for BackBone Assignments) has been 

demonstrated previously. 4-5 More recently we have examined projection 

experiments involving TOCSY or NOESY steps and aimed at side-chain 

assignments or structural studies. Combinations of projections from 

different experiments can be decomposed, optimized to a protein 

and experimental conditions. Thus, simultaneous decompositions of 

~50 projections from 5 different experiments on a histone provide 

15-dimensional data, where the advantages of many peaks in NOEbased 

dimensions are combined with the stronger signals in dimensions 

based on J-couplings. Time efficiency, flexible combination of projection 

sets and robustness characterise the PRODECOMP approach, opening 

up for a wide range of applications. PRODECOMP and SHABBA are 

being incorporated into the common software framework developed by 

the CCPN project 6 , and interfaced to instrument software (TopSpin) 7 . 

References: 

(1) Malmodin, D.; Billeter, M. Multiway Decomposition of NMR Spectra 

with Coupled Evolution Periods. J. Am. Chem. Soc. 127, 13486— 

13487 (2005). 

(2) Malmodin, D.; Billeter, M. Robust and versatile interpretation of 

spectra with coupled evolution periods using multi-way decomposition. 

Magn. Reson. Chem. 44, S185—S195 (2006). 

(3) Staykova, D.K.; Fredriksson, J.; Billeter M. PRODECOMPv3: 

Decompositions of NMR projections for protein backbone and sidechain 

assignments, and structural studies. Bioinformatics 24, 2258— 

2259 (2008). 

(4) Staykova, D.K.; Fredriksson, J.; Bermel W.; Billeter M. Assignment 

of protein NMR spectra based on projections, multi-way decomposition 

and a fast correlation approach. J. Biomol. NMR. 42, 87—97 (2008). 

(5) Billeter M., Staykova, D.K. Rapid Multidimensional NMR: 

Decomposition Methods and their Applications. Encyclopedia 

of Magnetic Resonance, John Wiley & Sons, Ltd. DOI: 

10.1002/9780470034590.emrstm1037 (2009). 

(6) Extend-NMR Project homepage, http://www.extend-nmr.eu/. 

(7) Bruker Biospin GmbH. 

So26 

Hairpin structure of a biarsenical-tetracysteine motif 

determined by NMR spectroscopy 

Madani, Fatemeh 1 ; Lind, Jesper 1 ; Damberg, Peter 1 ; Adams, Stephen R. 2 ; 

Tsien, Roger Y. 2 ; Gräslund, Astrid O. 1 

1 Stockholm University, Biochemistry and biophysics, Stockholm, 

Sweden; 2 University of California, San Diego, United States 

The biarsenical tetracysteine motif is a useful tag for genetic labeling of 

proteins with small molecules in living cells. The present study concerns 

the structure of a 12-amino acid peptide FLNCCPGCCMEP bound to the 

fluorophore ReAsH, based on resorufin. 1H-NMR was used to determine 

the solution structure of the complex formed between the peptide and 

the ReAsH moiety. Structure calculations based on the NMR results 

showed that the backbone structure of the peptide is fairly well defined 

with a hairpin-like turn, similar to a β turn type II, formed by the central 

CPGC segment. The most stable complex was formed when As2 was 

bonded to C4 and C5, and As1 was bonded to C8 and C9. Two clear 

NOESY crosspeaks between the Phe1 sidechain and ReAsH confirmed 

the close positioning of the phenyl ring of Phe1 and ReAsH. Phe1 was 

found to have an edge-face geometry relative to ReAsH. The close 

interaction between Phe1 and ReAsH may be highly significant for the 

fluorescence properties of the ReAsH complex. 


So27 

Selective detection of organophosphorus compounds with onflow 

heteronuclear correlation spectrometry in LC-NMR analysis 

Koskela, Harri 1 ; Ervasti, Mia 2 ; Björk, Heikki 1 ; Vanninen, Paula 1 

1 University of Helsinki, VERIFIN, Helsinki, Finland; 2 Helsinki Metropolia 

University of Applied Sciences, Technology and Transport, Helsinki, 

Finland 

Liquid chromatography hyphenated with nuclear magnetic resonance 

(LC-NMR) 1,2 is a useful technique in analysis of complex samples. 

However, the on-flow 1 H NMR spectrometry usually suffers from 

intense eluent peaks. The poor dynamic range can be improved either 

with use of deuterated eluents or with various signal suppression 

schemes. Deuterated eluents are expensive, and band-selective signal 

suppression schemes are often unsatisfactory in detection of chemicals 

at low concentration. 

If the analytes have a common heteronucleus, on-flow heteronuclear 

correlation spectrometry can offer several benefits. Here, we describe 

the use of on-flow one-dimensional proton-phosphorus heteronuclear 

single quantum coherence (1D 1 H- 31 P HSQC) spectrometry in selective 

detection of characteristic organophosphorus degradation products 

of nerve agents sarin and soman. 3 These chemicals were monitored 

during chromatographic separation at level 10 µg with high time 

resolution (1 spectrum per 2 s) with on-flow 1D 1 H- 31 P HSQC, while 

the eluent peaks and impurities were almost completely suppressed. In 

this way, the retention times of these non-UV-detectable analytes were 

determined for loop collection and SPE enrichment. The analytes, eluted 

from the SPE cartridges with deuterated solvent, were analyzed off-line 

using a micro coil probe head with high mass sensitivity. 4,5 

1. Albert, K. On-line LC-NMR and Related Techniques; Wiley: Chichester, 

2002. 

2. Keifer, P.A. Annu. Rep. NMR Spectrosc. 2007, 62, 1-47. 

3. Koskela, H.; Ervasti, M.; Björk, H.; Vanninen, P. Anal. Chem. 2009, 

81, 1262-1269. 

4. Schlotterbeck, G.; Ross, A.; Hochstrasser, R.; Senn, H.; Kühn, T.; 

Marek, D.; Schett, O. Anal. Chem. 2002, 74, 4464-4471. 

5. Koskela, H.; Vanninen, P. Anal. Chem. 2008, 80, 5556-5564. 

So28 

NMR study of hydrogen bonds geometries in homo-conjugates 

of carboxylic acids 

Guo, Jing 1 ; Tolstoy, Peter 1 ; Koeppe, Benjamin 1 ; Denisov, Gleb 2 ; Limbach, 

Hans-Heinrich 1 

1 Free University of Berlin, Berlin, Germany; 2 St.Petersburg State 

University, St.Petersburg, Russian Federation 

One of the major interests in hydrogen bond studies is to find out 

the position of the bridging proton. When crystallographic methods 

are not feasible one could rely on spectroscopic ones, which are 

valid if there are correlations between bridging proton position and 

spectroscopic observables, such as NMR chemical shifts.1,2 However, 

in many short (strong) OHO hydrogen bonds there is a fast proton 

tautomerism between donor and acceptor. In this case comparatively 

long-characteristic-time NMR measurements give parameters which 

are weighted averages of intrinsic values for individual tautomers. 

ABSTRACTS POSTER


In this work we focused our attention on intermolecular H-bonded 

complexes formed between a carboxylic acid and its carboxylate as 

well as symmetric intramolecular H-bonded hydrogen dicarboxylates. 

Key feature in these homo-conjugated complexes is that two proton 

tautomers are degenerated and contribute equally to the averaged 

spectroscopic values, which is to say that each average value is half 

the sum of two intrinsic ones. The compounds were dissolved in polar 

aprotic solvent (mixture of liquefied freonic gases) and studied by 1 H, 2 H 

and 13 C NMR spectroscopy at low-temperature down to 120 K. Analysis 

of the spectroscopic data allowed us to establish the correlation 

between H/D isotope effect on carboxylic carbon chemical shift and 

bridging proton position. Practical use of the correlation is that the 

geometric information about OHO hydrogen bonds could be read out 

directly from experimental 13 C NMR spectra. 

[1] P. M. Tolstoy, P. Schah-Mohammedi, S. N. Smirnov, N. S. Golubev, G. 

S. Denisov, H. H. Limbach, J. Am. Chem. Soc. 2004, 126, 5621-5634. 

[2] H. H. Limbach, P. M. Tolstoy, N. Pérez-Hernandez, J. Guo, I. G. 

Shenderovich, G. S. Denisov, Isr. J. Chem. Accepted. 

So29 

NMR investigation of the anticancer drug - Topotecan 

Hyz, Karolina 1 ; Bocian, Wojciech 2 ; Sitkowski, Jerzy 2 ; Bednarek, El¿bieta 2 ; 

Kawêcki, Robert 1 ; Kozerski, Lech 1 

1 Institute of Organic Chemistry, Polish Academy of Sciences, Warsaw, 

Poland; 2 National Medicines Institute, Warsaw, Poland 

Topotecan (TPT) is in clinical use as an antitumor agent, Hycamtin ® . It 

acts by binding to the covalent complex formed between nicked DNA 

and topoisomerase I, and inserts into the single-strand nick, thereby 

inhibiting the religation of the nick and acting as a poison. 

Topotecan structure in water at biological pH and its reactivity in 

different media has not been studied in details. Also, with regard to the 

interaction with DNA it is important to know the aggregation state and 

dynamics in solution of interacting molecules. 1 Here, we use NMR to 

trace its structure in aqueous medium, the tautomerism and solvolysis 

products in various solvents and conditions (CH 3 OH, DMSO, H 2 O). The 

reaction products of quaternization of nitrogen and methylation of 

oxygen sites are characterized by means of ESI MS and 1 H/ 13 C – HMBC 

and HSQCAD NMR. 2 We show that TPT tumbles in water solution as 

aggregate with the correlation time characteristic for large molecules. 

We have focused on the NMR characterization of the TPT with the 

anticipation that its aggregation, tumbling properties and intramolecular 

dipolar interactions will be a common feature for other compounds and 

can be useful in tracing the interactions of this class of topo I poisons 

with DNA. 

References 

1. W. Bocian, R. Kawêcki, E. Bednarek, J. Sitkowski, A. Pietrzyk, M. 

P. Williamson, P. E. Hansen, L. Kozerski, Chem. Eur. J., 2004, 10, 

5776-5787; W. Bocian, E. Bednarek, J. Sitkowski, R. Kawêcki, M.P. 

Williamson, P.E. Hansen , L. Kozerski, Chem. Eur. J., 2008, 14, 2788- 

2794; 

2. K. Hyz, W. Bocian, R. Kawêcki, E. Bednarek, J. Sitkowski, L. Kozerski, 

in preparation. 

118 

So30 

Stepwise progress towards the structure determination 

of a protein prone to self-association: the case of Human 

Papillomavirus E6 oncoprotein 

Zanier, Katia 1 ; Ould M’hamed Ould Sidi, Abdelahi 1 ; Rybin, Vladimir 2 ; 

Chapelle, Anne 1 ; Atkinson, Andrew 3 ; Kieffer, Bruno 3 ; Travé, Gilles 1 

1 Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch, France; 

2 EMBL, Heidelberg, Germany; 3 Institut de Génétique et de Biologie 

Moléculaire et Cellulaire, Illkirch, France 

Human Papillomavirus (HPV) E6 oncoproteins are responsible for various 

forms of epithelial cancers including cervical cancer. High-risk HPV E6 

oncoproteins interact with the ubiquitin ligase E6AP and target several 

cellular proteins, including p53 and several PDZ domain containing 

proteins, towards ubiquitin–mediated degradation. 

HPV E6 consists of two 75-residue zinc-binding domains (named 

E6-N and E6-C). We found that E6 proteins from different viral strains 

have in common the ability to self-associate into oligomeric as well as 

multimeric structures. Such property precludes the concentration of 

E6 samples required for structural studies. For this reason, structural 

studies initially focused on constructs of the isolated E6-N and E6-C 

domains. We observed that while the E6-C domain is monomeric 

even at high concentrations, the E6-N domain forms a weak dimer. 

Acquisition of NMR spectra at different concentrations allowed us 

to map the homodimer interface. By mutating hydrophobic residues 

situated at the interface, we obtained monomeric samples of the 

E6-N domain, ultimately enabling us to solve its structure by NMR 

spectroscopy. By introducing the same mutations into full-length E6, we 

were able to suppress polymerisation events and therefore to obtain the 

first NMR spectra of a full-length HPV E6 protein. It therefore appears 

that E6-N dimerization is the first of a series of events leading to fulllength 

E6 polymerization. Samples of monomeric full-length E6 were 

used to map the binding to E6AP derived peptides by chemical shift 

perturbation analysis. Structural, dynamic and interaction data on the 

monomeric E6 protein will be discussed. 

So31 

Revealing the mechanism of folding upon binding in 

intrinsically unfolded viral proteins using residual dipolar 

couplings and paramagnetic relaxation 

Jensen, Malene R. 1 ; Houben, Klaartje 1 ; Costanzo, Stéphanie 2 ; Lowry, 

David F. 3 ; Blanchard, Laurence 1 ; Marion, Dominique 1 ; Daughdrill, Gary 

W. 3 ; Ruigrok, Rob W. H. 4 ; Longhi, Sonia 2 ; Blackledge, Martin 1 

1 Institut de Biologie Structurale, Grenoble, France; 2 Bioénergétique et 

Ingénierie des Protéines, Marseille, France; 3 University of South Florida, 

Tampa, United States; 4 Unit for Virus Host Cell Interactions, Grenoble, 

France 

It is well known that protein structure and function are intertwined, 

and until recently it was generally thought that the native structures of 

functional proteins are folded or highly ordered. However, over the last 

decade it has become increasingly clear that a large fraction (~40%) of 

the proteins encoded by the human genome are intrinsically disordered, 

or contain unstructured regions of significant length (> 50 amino acids). 

These proteins play key roles in many physiological and pathological 

processes including signaling, cell cycle control, molecular recognition 

and transcription and replication. 

One of the most intriguing aspects of intrinsically disordered proteins 

(IDPs) is the capacity of many of these proteins to undergo structural 

transitions to folded forms upon binding to physiological partners. This 

disorder-to-order transition is a key feature of the complex relationship 

between the structural dynamics and function of intrinsically disordered 

proteins. Here, we study disorder-to-order transitions in nucleoproteins 

of Sendai virus (SeV) and Measles virus (MeV) – two members of the 


Paramyxoviridae family. In both viruses, replication and transcription 

of the viral RNA are initiated by an interaction between the intrinsically 

disordered C-terminal domain NTAIL of the nucleoprotein and the 

C-terminal three-helix bundle domain, PX, of the phosphoprotein P. 

Using chemical shifts, hetero-nuclear NOEs, residual dipolar couplings 

and paramagnetic relaxation we provide atomic resolution models of the 

disordered NTAIL proteins in their pre-recognition states and in complex 

with their partner proteins. The results provide intriguing insight into 

the forces controlling molecular recognition and reveal the importance 

of IDP-mediated fly-casting interactions in these highly dynamic 

complexes. 

So32 

Solution structure and dynamics of a psychrophilic thioldisulfide 

oxidoreductase 

Collins, Tony; Pais, Tiago M; Lamosa, Pedro; Santos, Helena; 

Matzapetakis, Manolis 

ITQB / UNL, Oeiras, Portugal 

Psychrophilic enzymes produced by cold-adapted micro-organisms 

have successfully overcome the low temperature challenge and have 

adapted to maintain high catalytic rates in their permanently cold 

environments [1]. The current consensus is that this high activity 

at low temperatures is mainly achieved through an increase in the 

flexibility of the protein structure, thereby allowing for the molecular 

motions necessary for activity in the low thermal energy environment. 

Nevertheless, while the demonstrated decreased stability and high 

activity at low temperatures of psychrophilic enzymes does support 

this hypothesis, there is, as yet, no direct experimental evidence of an 

increase in flexibility. In effect, the flexibility of a protein is a difficult 

parameter to assess since it may be related to the frequency or the 

amplitude of the fluctuations and limited to only a specific part of the 

protein, in particular at or near the active site. Previous attempts to 

demonstrate this proposed increased flexibility of psychrophilic enzymes 

used approaches such as measurement of hydrogen/deuterium 

exchange rates, fluorescence quenching, neutron scattering and even 

molecular dynamics simulation studies, with variable and conflicting 

results being obtained. We are using NMR to probe the flexibility of a 

thiol-disulfide oxidoreductase (DsbA) from an Antarctic bacterium as 

this technique offers the advantage of allowing analysis of both local 

motions and global movements over various time scales. The gene 

encoding the psychrophilic enzyme (187 amino acids) was isolated, 

cloned and overexpressed in E. coli and the 13 C 15 N-labeled protein 

was purified from the periplasmic extracts. Here, the results for the 

determination of the solution structure of the reduced form of this cold 

adapted oxidoreductase at pH 7.2 and 25 °C will be presented as well 

as the preliminary investigation of the flexibility of its structural edifice. 

[1] D’Amico, S., Collins, T., Marx, J.C., Feller, G. and Gerday, C. (2006) 

Psychrophilic microorganisms: challenges for life. EMBO Rep., 7, 385- 

389. 


So33 

Detection and characterisation by LC-SPE-NMR/MS of a new 

diasteromer of the aldehydic form of oleuropein aglycon in 

extra-virgin olive oil 

Pérez-Trujillo, Miriam 1 ; Gómez-Caravaca, Ana María 2 ; Segura-Carretero, 

Antonio 2 ; Parella, Teodor 1 

1 Universitat Autònoma de Barcelona, Servei de Ressonància Magnètica 

Nuclear, Bellaterra, Spain; 2 Universidad de Granada, Departamento de 

Química Analítica, Granada, Spain 

Many studies have demonstrated the important role of olive oil 

polyphenols as preventive agents against coronary heart disease and 

some cancers. A recent work has demonstrated the protective effects 

of one of the main olive oil polyphenols, oleuropein aglycon, against 

cell injury. 1 Until now, only two diasteromers of the aldehydic form of 

oleuropein aglycon have been identified in olive oil. 2 

In this work, the phenolic extract of an extra-virgin olive oil (Cornezuelo 

variety) was analysed by LC-SPE-NMR/MS. 3 These hyphenated 

techniques allowed the identification and complete 1 H and 13 C NMR 

characterisation of a new diasteromer of the aldehydic form of 

oleuropein aglycon. 

The HPLC analysis allowed the separation of the phenolic extract in 

more simple fractions. These fractions, initially detected by DAD and 

ESI-MS, were concentrated by 10 trapping processes in the SPE 

system. After that, they were transferred to the NMR cell and they 

were analysed by 1 H NMR experiments with double presaturation 

of the residual solvent signals. One of these fractions was studied 

more in detail. Its ESI(-)-MS spectrum showed a major peak at m/z 

377.2 corresponding to one or more isomers of oleuropein aglycon. 

A quick look of its 1 H NMR spectrum revealed four possible isomers. 

Several NMR experiments were performed in order to identify and 

fully characterised them, such as 1 H- 1 H COSY and TOCSY and 1 H- 13 C 

HSQC and HMBC experiments. The complete characterisation of them 

revealed the presence of a new diasteromer of the aldehydic form of 

oleuropein aglycon. 

1Paiva-Martins, F., Fernandes, J., Rocha, S., Nascimento, H., Vitorino, 

R., Amado, F., Borges, F., Belo, L., Santos-Silva, A. Mol. Nutr. Food Res. 

2009, 53, 609-601. 

2 Christophoridou, S., Dais, P. Anal. Chim. Acta 2009, 633, 283-292. 

3 Lindon, J. C., Nicholson, J. K., Wilson, I. D. Prog. NMR Spectrosc. 

1996, 29, 1-49. 

So34 

Variable temperature studies of the fluxional β-agostic silyl 

hydride complex (ArN)(η 2 -ArNSiMe 2 H)Mo(PMe 3 ) 2 Cl (Ar = 

2,6-dimethylphenyl) 

Rees, Nicholas 1 ; Mountford, Philip 1 ; Nikonov, Georgii 2 

1 University of Oxford, Chemistry Research Laboratory, Oxford, United 

Kingdom; 2 Brock University, Department of Chemistry, St Catharines, 

Canada 

Transition metal compounds with agostic Si-H…M interactions (1) have 

attracted significant recent attention, in part due to their relevance 

to a wide range of metal-mediated transformations of organosilicon 

compounds, such as hydrosilylation, dehydrogenative polymerization of 

silanes, silane alcoholysis. The β-agostic silyl hydride complex (ArN)(η 2 - 

ArNSiMe 2 H)Mo(PMe 3 ) 2 Cl (Ar = 2,6-dimethylphenyl) (2) is highly fluxional 

in solution, exhibiting four different exchange processes for the ligands 

attached to the metal center. Variable temperature proton and 31P NMR 

studies yield identical exchange rates for all these processes. This can 

be be explained by a highly correlated mechanism which proceeds via 

partial opening of the Si-H…Mo agostic bond, rotation around the N-Mo 

bond, and is completed by the ring closure on the other side of the 

complex. 

ABSTRACTS POSTER


1) (a) Kubas, G. J. “Metal Dihydrogen and σ-Bond Compoundes” Kluwer 

Academic/Plenum: New York 2001. (b) Crabtree, R. H. Angew. Chem. 

Int. Ed. Engl. 1993, 32, 789. (c) Corey, J. Y.; Braddock-Wilking, J. Chem. 

Rev., 1999, 99, 175. (d) Lin, Z. Chem. Soc. Rev. 2002, 31, 239. (e) 

Nikonov, G. I. Adv. Organomet. Chem., 2005, 53, 217. 

2) Ignatov, S. K.; Khalimon, A. Y. ; Rees, N. H. ; Razuvaev, A. G. ; 

Mountford, P. ; Nikonov, G. I. Inorg. Chem. in Press 

So35 

Comparison of the techniques for determining sulfonation 

degree of SPEEK by 1H-NMR, TGA and titration methods for 

PEMFC applications 

Turdu, Nursen 1 ; E. Unveren, Elif 1 ; Y. Inana, Tulay 1 ; Birkan, Burak 2 

1 TUBITAK, Marmara Research Center, Chemistry Institute, Kocaeli, 

Turkey; 2 Türk Demirdöküm Fabrikalarý A.Þ, Bozüyük/ Bilecik, Turkey 

Polymer electrolyte membrane (PEM) acts as a proton exchange 

membrane by transferring protons from anode to cathode in PEM fuel 

cell. NafionTM which is the most common used as a polymer electrolyte 

in PEMFC has high proton conductivity, good chemical stability but it 

has certain drawbacks such as cost and high fuel permeability. In order 

to overcome these drawbacks, there is an increasing demand to create 

novel polymeric materials with improved properties such as proton 

conductivity, fuel permeability and cost1. 

Poly(ether ether) ketone (PEEK) is a high performance engineering 

thermoplastic which is well known for having excellent mechanical 

properties, high thermo-oxidative resistance and also stability under 

acidic conditions. Therefore, sulfonated PEEK (SPEEK) is a potential 

material for polymer electrolyte fuel cells2. In this work, commercially 

available PEEK was sulfonated by using concentrated sulfuric acid 

at 60°C and SPEEK having different sulfonation degree values were 

obtained. 

The main objective of this work was to compare 1H NMR (500 MHz, 

Bruker spectrometer) TGA (Perkin Elmer, Pyris 1) and also titration 

methods for determining sulfonation degree of sulfonated polymers. The 

1H NMR (500 MHz) spectra were recorded at room temperature. 

As a result, NMR technique among these three methods was found to 

be quick and valuable technique for determination of DS. 

References 

1. M.A. Hickner, H. Ghassemi, Y.S. Kim, B.R. Einsla, J.E. McGrath, 

Chemical Reviews 104 (2004) 4587 – 4612. 

2. P. Xing, G. P. Robertson, M. D. Guiver, S. D. Mikhailenko, K. Wang, S. 

Kaliaguine, J Membr Science 229 (2004) 95-106. 

So36 

Exploitation of bile acids and their derivatives in synthesis of 

salophen-metal complexes 

Jurček, Ondřej 1 ; Cametti, Massimo 1 ; Kolehmainen, Erkki 1 ; Rissanen, 

Kari 1 ; Wimmer, Zdeněk 2 ; Drašar, Pavel 3 

1 University of Jyväskylä, Department of Chemistry, Jyväskylä, Finland; 

2 Institute of Experimental Botany AS CR, Isotope laboratory, Prague, 

Czech Republic; 3 Institute of Chemical Technology Prague, Department 

of Chemistry of Natural Compounds, Prague, Czech Republic 

Bile acids are wedge-shaped, amphipathic C24-molecules, 

predominantly found in the bile of mammals. They play important role in 

many biological functions [1]. On the other hand, salophens are widely 

used tetradentate ligands and their stable complexes with metals or 

actinides have been successfully employed as receptors, catalysts, and 

carriers [2]. Merging these two structural units into one single entity 

provides a novel class of the metal complexes, which may possess 

biological activity, and interesting properties such as transport through 

120 

membranes and recognition of neutral and negatively charged biological 

target species. 

Different ligands containing lithocholyl, dehydrocholyl or cholyl moieties 

or their derivatives were synthesized and characterized by NMR and ESI 

TOF MS techniques. Preparation, isolation and characterization of their 

complexes with different metals are subject of our current interest. 

[1] a) Jenkins, G.; Hardie, L. J. (Eds.), Bile Acids: Toxicology and 

Bioactivity, RSC Publishing, 2008, p. 6; b) Hofmann, A.F.; Hagey, R. L., 

Cell. Mol. Life Sci., 2008, 65, 2461-2483; c) Virtanen, E.; Kolehmainen, 

E., Eur. J. Org. Chem. 2004, 16, 3385-3399. 

[2] a) Vigato, P. A.; Tamburini, S., Coord. Chem. Rev. 2004, 248, 

1717-2128; b) Cametti, M.; Nissinen, M.; Dalla Cort, A.; Mandolini, L.; 

Rissanen, K., J. Am. Chem. Soc. 2005, 127, 3831-3837; c) Cametti, 

M.; Nissinen, M.; Dalla Cort, A.; Mandolini, L.; Rissanen, K., J. Am. 

Chem. Soc. 2007, 129, 3641-3648; d) Sessler J.L.; Melfi P.J.; Pantos 

G.D., Coord. Chem. Rev. 2006, 250, 816-843. 

A financial support through the projects 127006 (decision date 

December 12th 2008) (O.J., E.K.) of the Academy of Finland, 2B06024 

(SUPRAFYT) (O.J., Z.W.) and MSM6046137305 (O.J., P.D.) of MŠMT ÈR 

are gratefully acknowledged. 

So37 

Design of combined probes for NMR/UV-Vis and CIDNP/LFP 

measurements 

Tolstoy, Peter 1 ; Koeppe, Benjamin 1 ; Ivanov, Konstantin 2 ; Yurkovskaya, 

Alexandra 2 ; Vieth, Hans-Martin 2 ; Limbach, Hans-Heinrich 1 

1 Free University of Berlin, Institute of Chemistry and Biochemistry, Berlin, 

Germany; 2 Free University of Berlin, Institute of Physics, Berlin, Germany 

The choice of a spectroscopic method limits the set of accessible 

structural and dynamic parameters for a given system. As NMR has a 

low time resolution, it is if often needed to couple it a technique with 

a shorter “time-scale”. To ensure the compatibility of spectra obtained 

by two methods it is also desired to perform the measurements on 

the same sample. As a possible solution, we propose the design of 

combined probes for NMR/UV-Vis and CIDNP/Laser Flash Photolysis 

(FLP) measurements. 

The liquid-state NMR/UV-Vis probe has an additional channel for fiberoptical 

probe placed below the NMR sample. The UV-Vis absorption 

spectra are measured in reflection from the PTFE insert. Short optical 

path length allows one to work with the highly concentrated samples 

suitable for the standard NMR measurements. We present and discuss 

the application of this setup for the investigation of the H-bonded 

complexes. [1] 

The CIDNP/LFP probe is an NMR probe augmented by three quartz light 

guides. One of the light guides is used for the sample irradiation by 

the laser flash, while two others are used for the transient absorption 

measurements. We present the results of the test measurements 

performed outside the NMR magnet and show that using this setup 

it is possible to get direct information on radical formation and decay 

kinetics. This information allows one to analyze CIDNP spectra more 

straightforward and reliable, without the necessity to vary the initial 

concentrations of the reactants. 

[1] P. M. Tolstoy, B. Koeppe, et al, Angew. Chem. 2009, DOI: 10.1002/ 

anie.200806181. 


So38 

Low-power-composite-CPMG G-BIRD adiabatic HSQMBC, an 

improved sequence for determination of heteronuclear longrange 

coupling constants 

Boros, Sándor 1 ; E. Kövér, Katalin 2 

1 Sanofi-aventis / Chinoin Co., Discovery Analytics, Budapest, Hungary; 

2 University of Debrecen, Department of Inorganic and Analytical 

Chemistry, Debrecen, Hungary 

Heteronuclear single-quantum multiple bond correlation (HSQMBC) 

experiment has been proven to be a very powerful method for the 

measurement of heteronuclear long-range coupling constants. The 

experiment provides the desired heteronuclear couplings for both 

quaternary and protonated carbons which can be extracted from the 

antiphase multiplets in the 1H dimension. 

Our poster demonstrates a variant of HSQMBC which contains an 

advantageous combination of the modifications available in the 

literature. New approach in this version is that the CPMG decoupling 

is applied on low power in order to avoid the unwanted heating of the 

sample and the probehead. A modified position is suggested for the 

blanking of gradient command in order to avoid the eddy-current in the 

detector coil. 

The performance and the robustness of the proposed experiment was 

tested with respect to different experimental parameters. The selective 

1D version of the method is also presented. 

References: 

R. T. Williamson, A. Boulanger, A. Vulpanovici, M. A. Roberts, W. H. 

Gerwick: J. Org. Chem., 67, 7927-7936 (2002) 

M. Köck, R. Kerssebaum, W. Bermel: Magn. Reson. Chem., 41, 65-69 

(2003) 

V. Lacerda J., G. V. J. da Silva, M. G. Constantino, C. F. Tormena, R. T. 

Williamson, B. L. Márquez: Magn. Reson. Chem., 44, 95-98 (2006) 

K. E. Kövér, Gy. Batta, K. Fehér: J. Magn. Reson., 181, 89-97 (2006) 

So39 

Structure and dynamics of benzodiazacoronads in the liquid 

phase - attempt to study of preorganization mechanism leading 

to planar chirality of crystals 

Kazmierski, Slawomir 1 ; Jurczak, Janusz 2 ; Sobczuk, Adam 2 ; Kalisiak, 

Jaroslaw 2 ; Nowicka, Katarzyna 1 ; Potrzebowski, Marek 1 

1 Centre of Molecular and Macromolecular Studies Polish Academy of 

Sciences, NMR Laboratory, Lodz, Poland; 2 Institute of Organic Chemistry 

Polish Academy of Sciences, Warszawa, Poland 

Chiral crystals formed from achiral molecules have recently received 

a great deal of attention due to their attractive structural properties 

and prospective applications in chemistry[1]. For instance, such 

compounds can be used as ligands in enantioselective reactions or 

models for investigation of molecular recognition[2].As a part of our 

interest in the problem of formation of crystals with planar chirality 

[4], we have presented the structural studies for two diazacoronands 

1 and 2 [3] in the solid phase. It was apparent, that crystals of 1 and 

2 have to be considered as a two-component system consisting of 

an organic unit and a water molecule in 1:1 ratio. Both components 

play an important role in the crystal structure. The strong (O-H O, N-H 

O) and weak (C-H O) intermolecular hydrogen bonds are responsible 

for phase organization and - in consequence - formation of chiral or 

achiral crystals. The alignment of the water molecule with respect to the 

macrocycle is different for samples 1 and 2, so the water molecule can 

be an important achiral cofactor responsible for chiral crystallization. 

It is worthy to note, that process of removal of water from the crystal 

lattice of 1 is reversible.In this work, the results of the investigation 

of the structure and influence of the water on the dynamics of two 

benzodiazacoronads 1 and 2 in the liquid phase will be presented. 

The major aim of our project is to understand the origin of distinction 

between these compounds and answer the questions: 

- how does the water molecule alter the intramolecular dynamics of the 

molecules under investigation? 

- what is the role of water in the formation process of chiral crystals 

during the preorganization step? 

The results of the NMR relaxation parameters measurements will be 

presented and discussed. 

1Kondepudi, D. K.; Kaufman, R. J.; Singht, N. Science 1990, 250 

2Dahmen, S.; Brase, S. J. Am. Chem. Soc. 2002, 124, 5940;Gibson, S. 

E.; Knight, J. D. Org. Biomol.Chem.2003, 1, 1256 

3Kalisiak, J.; Jurczak, J. Synlett 2004, 9, 1616-1618; Kalisiak, J.; 

Jurczak, J. Cryst. Growth Des. 2006, 6, 22-24 

4Pacholczyk, J.; Kalisiak, J.; Jurczak, J.; Potrzebowski M.J. J. Phys. 

Chem. B 2007, 111, 2790-2799 


So40 

Structural polymorphism of 441-residue tau at single residue 

resolution 

Bibow, Stefan 1 ; Mukrasch, Marco D. 1 ; Korukottu, Jegannath 1 ; 

Jeganathan, Sadasivam 2 ; Biernat, Jacek 2 ; Griesinger, Christian 1 ; 

Mandelkow, Eckhard 2 ; Zweckstetter, Markus 1 

1 MPI Biophysical Chemistry, NMR Based Structural Biology, Goettingen, 

Germany; 2 Max Planck Unit for Structural Molecular Biology, c/o DESY, 

Hamburg, Germany 

Alzheimer’s disease is characterized by abnormal protein deposits 

in the brain, such as extracellular amyloid plaques and intracellular 

neurofibrillary tangles. The tangles are made of a protein called tau 

comprising 441 residues in its longest isoform. Tau belongs to the class 

of intrinsically disordered proteins, binds to and stabilizes microtubules 

and partially folds into an ordered β-structure during aggregation 

to Alzheimer paired helical filaments (PHFs). Here we show that it 

is possible to overcome the size limitations that have traditionally 

hampered detailed nuclear magnetic resonance (NMR) spectroscopy 

studies of such large non-globular proteins. This is achieved using 

optimal NMR pulse sequences and matching of chemical shifts from 

smaller segments in a divide and conquer strategy. The methodology 

reveals that 441-residue tau is highly dynamic in solution with a distinct 

domain character and an intricate network of transient long-range 

contacts important for pathogenic aggregation. Our results establish 

that NMR spectroscopy can provide detailed insight into the structural 

polymorphism of very large nonglobular proteins. 

So41 

Dynamics of the lid-segment control Hsp90 ATP-turnover 

Lagleder, Stephan; Hagn, Franz; Buchner, Johannes; Kessler, Horst 

Technische Universität München, Chemistry, Garching, Germany 

Hsp90 is a molecular chaperone which plays a key role for the 

activation of numerous client proteins. This process is coupled to the 

binding and hydrolysis of ATP in its N-terminal domain. An important 

structural element is the Hsp90 ATP-lid, a segment which closes 

the ATP-binding pocket after nucleotide binding. Deletion of the lidsegment 

facilitates N-terminal dimerization of Hsp90 but abolishes 

ATP-hydrolysis. Thus the lid-segment seems to be fundamental for the 

function of Hsp90. The interaction between helix 1 and the ATP-lid in 

the Hsp90 N-terminal domain has been shown to play an important role 

for the flexibility of the lid-segment. We analyzed how the lid-dynamics 

influence the Hsp90 ATPase and chaperone activity. We show that a 

mutation within the ATP-lid that weakens its interaction with helix 1 

ABSTRACTS POSTER


slightly, leads to a several-fold increase in the Hsp90 ATP-turnover and 

impairs its in vivo function at elevated temperatures. On the other hand, 

the mutation of two hydrophobic residues in helix 1 confers a much 

higher degree of flexibility to the ATP-lid. But, surprisingly, in this case 

the increased lid-dynamics do not result in increased Hsp90 ATPaseactivity. 

On the contrary, the ATP-turnover is significantly reduced. These 

results indicate that the dynamics of the ATP-lid are precisely tuned to 

ensure Hsp90 ATP-turnover at a physiologically appropriate rate. 

So42 

Improving resonance assignment of intrinsically unstructured 

proteins 

Dürr, Ulrich H. N.; Xiang, Sheng-Qi; Narayanan, Raghav L.; Zweckstetter, 

Markus 

Max-Planck-Institute for biophysical chemistry, NMR-based structural 

biology, Göttingen, Germany 

The outstanding role of intrinsically unstructured proteins in many 

biological contexts is now widely appreciated [1]. Similarly, protein 

states that are actively unfolded by denaturing agents can be helpful to 

gain insight on functional states. NMR spectroscopy is the method of 

choice to study dynamic and conformational properties of such proteins. 

We have investigated how unstructured proteins behave in innovate 

NMR spectroscopic schemes that have previously been applied to 

folded proteins. 

The BEST (band selective excitation short transient)-NMR scheme [2] 

increases spectral sensitivity by rapid pulsing. Our experiments show 

that BEST-experiments can successfully be applied to intrinsically 

unstructured -synuclein and urea-unfolded ubiquitin, but sensitivity 

improvement is not as convincing as it is for folded ubiquitin. 

Automated projection spectroscopy (APSY, ref. 3) could easily resolve 

the heavily overlapped resonances of sizable unfolded proteins. We 

implemented a BEST-version of a six-dimensional APSY-experiment. 

Moreover, we tested different solvent suppression schemes to 

replace pre-saturation which is not applicable to unfolded proteins. 

The presence of highly concentrated urea as denaturing agent poses 

additional challenges, especially for variable temperature experiments 

on cryogenic probes. The mentioned technical problems could be 

overcome, and sequential information was gained from six- and sevendimensional 

APSY spectra. We demonstrate that the program MARS 

[4], which was developed in our laboratory, is an ideal choice to analyze 

APSY-generated peak lists. MARS was used to automatically assemble 

almost complete assignments from APSY-data. 

References 

[1] Dyson and Wright, Nat Rev Mol Cell Biol 6, 197. 

[2] Schanda and Brutscher, JACS 127, 8014, Lescop et al., JMR 

187,163. 

[3] Hiller et al., PNAS 102, 10876. 

[4] Jung and Zweckstetter, JBNMR 30, 11. 

So43 

NMR study of transmembrane domain of pro-apoptotic protein 

BNip3 

Pustovalova, Yulia; Bocharov, Eduard; Schulga, Alexey; Goncharuk, 

Marina; Volynsky, Pavel; Arseniev, Alexander 

Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, 


Mitochondrial pro-apoptotic protein BNip3 plays an important role in 

hypoxia-induced death of normal and malignant cells. BNip3 belongs 

to BH3-only subfamily of Bcl-2 family but it has atypical mechanism of 

activity. Cell death mediated by BNip3 is independent of cytochrome 

122 

c release and shows several characteristics of necrosis. Both the 

pro-apoptotic activity and BNip3 interaction with other Bcl-2 family 

members depends on the C-terminal transmembrane (TM) domain, 

which dimerizes tightly in lipid environments and this association has 

strong sequence dependence. The right-handed parallel helix-helix 

structure and internal dynamic properties of the dimeric TM fragment 

146-190 of human BNip3 embedded in lipid DMPC/DHPC bicelle was 

determined by solution heteronuclear NMR spectroscopy. The unique 

structure of the dimeric domain with hydrogen bond rich His-Ser node 

in the middle of membrane, accessibility of the node for water, and 

continuous hydrophilic track across the membrane suggest that the 

domain can provide an ion conducting pathway through the membrane. 

Indeed, the BNip3 TM domain was shown to induce conductivity of 

artificial bilayer lipid membrane in a pH-dependent and potassium 

independent manner that can be considered as circumstantial evidence 

of proton selective conduction through the dimer. These findings and 

currently available information about phenomenology of programmed 

cell death allow us to propose a mechanism of triggering necrosis-like 

cell death by BNip3 in case of hypoxia-acidosis of human tissues. For 

further investigation we developed system of bacterial expression and 

purification of full-length BNip3 and acquired preliminary NMR spectra 

of the full-length protein in membrane mimicking environment. 

So44 

Optimization of the NMR spectrum of the 37 kDa enzyme 

NADPH: Protochlorophyllide Oxidoreductase 

Proudfoot, Andrew; Hunter, C Niel; Craven, C Jeremy; Williamson, Mike 

P 

University of Sheffield, Dept of Molecular Biology and Biology, Sheffield, 


The light-driven enzyme protochlorophyllide oxidoreductase (POR) is 

responsible for catalysing the reduction of the C 17 - C 18 double bond of 

the D ring of protochlorophyllide (Pchlide) in the presence of NADPH, 

forming chlorophyllide (Chlide). The reaction catalysed by POR is a key 

step in chlorophyll biosynthesis and is essential in the development of 

chloroplasts. Due to POR being a light-activated enzyme, the enzymesubstrate 

complex can be pre-formed in the dark prior to the initiation 

of catalysis. This characteristic coupled to the use of NMR is expected 

to lead to a greater understanding of the hydride transfer reaction, and 

of the role of internal dynamics in the function of POR. 

We describe studies aimed at improving the yield, stability and 

spectrum of 13 C, 15 N, 2 H triple-labelled POR. Induction of expression 

using IPTG was shown to produce protein expression for 2.5 hours 

following the addition of IPTG, whatever the growth stage, leading 

to improved yield by adding the IPTG at high cell density. To assess 

suitable solution conditions, thermofluor experiments were conducted. 

These experiments observe the change in fluorescence of SYPRO 

Orange, in the presence of a range of salts and pH values, as a function 

of temperature. [SYPRO Orange fluoresces upon binding to hydrophobic 

regions, indicating thermal unfolding.] Stability was shown to depend 

crucially on purification conditions and rigorous use of protease 

inhibitors at all stages. Back exchange of some backbone HN protons 

following growth in D 2 O is extremely slow, hindering their observation. 

Combined temperature and pH control has been used to produce 

conditions under which almost all of the 322 expected HSQC signals 

can be observed. 


Solid State Physics 

Sp10 

Single-Crystal NMR for the layered semiconductor TlGaSe2 

Panich, Alexander 1 ; Kashida, Shoji 2 

1 Ben-Gurion University of the Negev, Physics, Beer Sheva, Israel; 

2 Niigata University, Department of Environmental Science, Niigata, Japan 

We report on 69Ga and 205Tl NMR study of the single crystal of 

thallium gallium selenide [1]. Our findings show that transformation 

from the high temperature paraelectric phase to the low temperature 

ferroelectric phase occurs via an incommensurate phase that exists in 

the temperature range from Tc = 107.5 to Ti = 118 K. On approaching 

phase transition at Ti from above, the crystal exhibits a soft mode 

behavior, which is somewhat different for thallium and gallium 

substructures. Redistribution of 69Ga line intensities with temperature 

in the ferroelectric phase indicates a variation of the domain structure 

of this phase. 

[1] A. M. Panich, S. Kashida, J. Phys.: Condens. Matter 20(39) 

395211/1-8 (2008). 

Sp11 

35 Cl spin-lattice relaxation in mercuric chloride 

Keartland, Jonathan 

University of the Witwatersrand, School of Physics, Johannesburg, South 

Africa 

Measurements of the 35 Cl spin-lattice relaxation rates in the molecular 

solid HgCl 2 have been made in the temperature range 10 K - 480 K 

using a coherent pulsed nuclear quadrupole resonance spectrometer 

operating in the range 19 - 23 MHz. Various variable temperature 

arrangements were used, as well as fixed temperature baths. The 

standard inversion-recovery three pulse sequence was used to 

monitor the evolution of the 35 Cl quadrupolar spin-echo as a function 

of the spacing between the inversion pulse and the echo sequence. 

The results (up to a temperature of approximately 400K) support 

the supposition that nuclear spin-lattice relaxation in this material is 

dominated by interactions of the nuclear quadrupole moment with 

molecular librations and/or phonons, as might be expected in an 

insulating solid of this type. If a single mode Einstein model is used, 

a mode of approximately 30 cm —1 is responsible for spin-lattice 

relaxation. If a two-phonon Raman model is assumed, then a Debye 

temperature of approximately 50 K is extracted from the results. At 

temperatures above 400 K there are deviations from the behaviour 

predicted by the theoretical models, and it is suggested that these are 

evidence of either pre-melting effects (the melting temperature of HgCl 2 

is 550 K), or molecular re-orientations. 

In addition to the spin-lattice relaxation measurements, the quadrupolar 

linewidth was measured over the entire temperature range. At this stage 

the results are poorly understood, but it would appear that the linewidth 

decreases steadily at as the temperature increases at temperatures 

below 300 K, reaches a minimum around room temperature and then 

increases steadliy thereafter. 

Details of the results, and the models used in data analysis will be 

presented. 


Sp12 

Orientation-dependent NMR study of the Y-Al 76 Co 22 Ni 2 complex 

metallic alloy 

Vrtnik, Stanislav; Dolinšek, Janez; Jeglič, Peter 

J. Stefan Institute, Condensed Matter Physics, Ljubljana, Slovenia 

We present a 27 Al NMR angular-dependent study of the Y-Al 76 Co 22 Ni 2 

complex metallic alloy. Alloy has monoclinic unit cell with lattice 

parameters a=1,7071 nm, b=0,4099 nm, c=0,7491 nm, β=116,7° 

and 32 atoms in the unit cell, which are placed on 9 crystalgraphically 

inequivalent atomic positions (2 Co/Ni and 7 Al). Our single crystal 

was grown by the Czochralski method using a native seed and cut into 

three bar-shaped samples (2x2x6 mm 3 ), with their long axes along 

three orthogonal direction: [010], [001] and direction perpendicular 

to the [010]−[001] plane. The so-prepared samples enabled us to 

perform crystallographic-direction-dependent NMR studies. The angular 

dependence of the NMR lineshapes were measured in magnetic field 

of 9,4 T at 80 K with axes of rotation parallel to the long side of the 

samples and perpendicular to the magnetic field. Angular dependent 

spectra exhibit central-transition peaks belonging to inequivalent 

aluminum atoms. The positions of peaks in the spectrum are changing 

with rotation of the samples and show patterns characteristic for Knight 

shift anisotropy and electric quadrupole interaction. Lines are wide and 

they overlap but we extracted the Knight shift and EFG tensors for two 

Al sites with the largest quadrupole coupling constant. 

Sp13 

New type of EPR response in dense intermetallic systems 

Ivanshin, Vladimir 1 ; Litvinova, Tatyana 1 ; Sukhanov, Andrey 2 

1 Kazan State University, MRS Laborastory, Kazan, Russian Federation; 

2 Kazan Physical-Technical Institute, Kazan, Russian Federation 

In typical EPR setups the host metallic ions in the undoped intermetallic 

systems are not an approptiate EPR probes. The usual spin fluctuation 

rate of these ions causes a huge EPR linewidth which makes the 

EPR signal undetectable. It is necessary to dope small amounts of 

paramagnetic ions with localized magnetic moments into the system 

under investigation in order to yield any measurable EPR signal. A 

rarther narrow Yb 3+ EPR in a dense heavy fermion compound YbRh 2 Si 2 

was totally unexpected [1]. Recent theoretical studies [2, 3] indicate 

that the hybridization of the 4f- and itinerat electrons, Fermi-liquid 

interaction, and short-range ferromagnetic correlations between 

the localized spins can lead to observation of narrow EPR signals in 

certain undoped Kondo-lattice compounds. Hence, it is expected that 

the strength of hybridization strongly influences the EPR behavior 

of the material. We have detected and studied the EPR in several 

intermetallics, such as YbRh 2 Pb, YbBiPt, and YbT 2 Zn 20 (T= Co, Fe) [4]. 

Temperature dependent EPR spectra reveal an anomalous duality of 4f 

and 3d electrons. New mechanism of the formation of EPR response 

based on the hybridization effects between localized and conduction 

electrons is proposed. 

1. J. Sichelschmidt, V.A. Ivanshin et al., Phys. Rev. Lett. 91, 156401 

(2003). 

2. E. Abrahams, P. Wölfle, Phys. Rev. B 78, 104423 (2008). 

3. P. Schlottmann, Phys. Rev. B 79, 045104 (2009). 

4. V.A. Ivanshin et al., J. Alloys. Compd. (2008), in press, doi:10.1016/j. 

jallcom.2008.09.172. 

ABSTRACTS POSTER


Sp14 

Photo-induced ions radicals formed in solid films of C60, PCBM, 

C120-O and C120-O-PCBM composites blended with M3EH- 

PPV polymer. ESR X, K- bands spectroscopy and relaxation 

parameters at 77 and 125 K 

Konkin, Alexander 1 ; Ritter, Uwe 1 ; Scharff, Peter 1 ; Aganov, Albert 2 ; Egbe, 

D.A.M. 3 ; Sariciftci, Serdar 3 

1 Ilmenau Technical University, Ilmenau, Germany; 2 Kazan State 

University, Kazan, Russian Federation; 3 LIOS, Linz, Austria 

Light-induced cation/anion radicals in solid thin films of M3EH-PPV 

polymers with mono-fullerenes C60 and PCBM as well as with 

di-fullerenes C120-O and C120-O-PCBM have been studied by 

means of ESR at 77K under Xe-lamp illumination. The g-factors of 

the positive polaron ESR spectra of M3EH-PPV obtained by the X,Kbands 

technique at 77K 125K respectively in all blends are around 

(± 0.0002) of g x = 2.0036, g y = 2.0025, g z = 2.0024 . It was found 

that g- factor components of the light-induced anion radicals (R LI ) 

registered in the blends with di-fullerenes C120-O and C120-O-PCBM 

do not correspond to the g-factor of anion radicals “dark” spectra of 

(C120-0) − and (C120-0) 2− di-fullerenes obtained electrochemically 

(were reported before in literature, g iso = 2.003) and “dark” spectra 

of original C120-O and C120-O-PCBM compound (powder) reported 

here (g iso = 2.003). Regarding the comparison of T 2 ~10 -8 for the mixed 

composites, they are similar to P3HT/PCBM as well as T 1 of R LI anion 

radicals approximately near the one order of magnitude shorter than T 1 

of M3EH-PPV polaron. 

Sp15 

NMR/NQR investigation of superconductive ferropnictides 

Dalibor, Paar 1 ; Grafe, Hans-Joachim 1 ; Lang, Guillaume 1 ; Hammerath, 

Franziska 1 ; Manthey, Katarina 1 ; Wolter, Anja 1 ; Curro, Nick J. 2 ; Büchner, 

Bernd 1 

1 IFW Dresden, Institute for Solid State Research, Dresden, Germany; 

2 University of California, Department of Physics, Davis, CA, United States 

The recent discovery of superconductivity in the layered ferropnictides 

RO 1-x F x FeAs (R=rare earth) has raised great interest within the solid 

state community. We have performed 139La, 57Fe, and 75As NMR 

and NQR measurements on the LaO 1-x F x FeAs oriented powders at 

temperatures up to 480 K. For all three nuclei in the x=0.1 material, it is 

found that the local Knight shift increases monotonically with increasing 

temperature, and scales with the macroscopic susceptibility, suggesting 

a single magnetic degree of freedom. The spin lattice relaxation rate 

for all nuclei also scale with one another. This result suggests a lack of 

any q-space structure in the dynamical spin susceptibility that might be 

expected in the presence of antiferromagnetic correlations. Rather, our 

results are more compatible with simple quasi-particle scattering. 

Our measurements reveal a strong anisotropy of the spin lattice 

relaxation rate, which suggest that superconducting vortices contribute 

to the relaxation rate when the field is parallel to the c-axis but not 

for the perpendicular direction. In the superconducting state, we find 

evidence for line nodes in the superconducting gap and spin-singlet 

pairing. 

This presentation was supported by FP7-REGPOT 229390 (Solid state 

NMR laboratory, Zagreb). 

[1] Grafe H-J et al., New J. Phys. 11 (2009) 035002 

[2] Grafe H-J et al., Phys. Rev. Lett. 101 (2008) 047003 

124 

Transport and Diffusion 

Td10 

T 1 -DOSY: Analysis of diffusion-relaxation NMR data with 

PARAFAC 

Botana Alcalde, Adolfo; Nilsson, Mathias; Morris, Gareth 

University of Manchester, School of Chemistry, Manchester, United 

Kingdom 

DOSY (Diffusion-Ordered Spectroscopy) 1 is one of the most commonlyemployed 

methods for identifying compounds in mixtures by NMR. 

However, this technique struggles to resolve the component spectra 

when there is severe signal overlap and/or the diffusion coefficients 

are very similar. In order to improve resolving power, T 1 relaxation 

was incorporated into diffusion experiments as a further dimension. 

This results, to a first approximation, in a trilinear dataset which, in 

contrast with a bilinear dataset (e.g. a standard DOSY dataset), can be 

decomposed with multivariate statistical methods such as PARAFAC 

(Parallel Factor Analysis) 2 to obtain physically correct component 

data (spectrum, diffusional decay, and relaxation evolution for each 

component). 

A mixture of simple alcohols with overlapping NMR signals was chosen 

as a test sample to assess this experiment, because of the relatively 

similar diffusion coefficients and spread in relaxation rates . A variety 

of pulse sequences derived from Oneshot 3 were developed for the 

acquisition of the 3D T 1 -DOSY data. The individual spectra from the 

mixture components were extracted by analyzing a number of spectral 

windows, containing signal overlap, independently. These windows were 

chosen as to maximize the trilinearity (i.e. only one multiplet component 

from each mixture component per window). This new method shows 

very promising results but it is worth noting that cross-correlated 

relaxation can cause deviations from strict trilinearity, leading to the 

spectral components obtained with PARAFAC showing cross-talk. 

1. “Diffusion-Ordered Spectroscopy”, G.A. Morris, in Encyclopedia of 

Magnetic Resonance: Volume 9, ed. D.M. Grant and R.K. Harris, J. Wiley 

and Sons, Ltd., 35-44 (2002). 

2. “PARAFAC. Tutorial and applications”, R. Bro, Chemometrics and 

Intelligent Laboratory Systems, 38:149– 171 (1997). 

3. “A One-Shot Sequence for High Resolution Diffusion Ordered 

Spectroscopy”, M.D. Pelta, G.A. Morris, M.J. Stchedroff and S.J. 

Hammond, Magnetic Resonance in Chemistry, 40, 147-152 (2002). 

Td11 

PFG-NMR and Goldman-Shen study of water diffusion and 

cross-relaxation in polyelectrolyte multilayers 

Wende, Christina; Schönhoff, Monika 

Institute of Physical Chemistry, Westfälische Wilhelms-Universität, 

Münster, Germany 

The self-assembly of polyelectrolytes of alternating charge from 

aqueous solutions onto charged surfaces leads to the formation of 

multilayered films. Their permeation properties for different types 

of molecules and their porosity are of general interest [1-3] , since the 

multilayers can act as separation membranes or colloidal hollow 

carriers. A direct determination of diffusion coefficients of small 

molecules within and through the multilayers is attractive, but hard to 

achieve. 

Here, we study stacked free-standing films of poly(styrene sulfonate)/ 

poly(diallyldimethyl ammoniumchoride), which are equilibrated at 

different relative humidities. The diffusion of water molecules is 

investigated by means of Pulsed-Field-Gradient-NMR. A non-Gaussian 


diffusion behaviour is found, which strongly depends on the relative 

humidity. Furthermore, a pronounced dependence of the PFG echo 

decay on the diffusion time is observed, which led to an evaluation in a 

model of restricted diffusion in a porous structure. 

In addition the influence of magnetization exchange between water 

and polymer spins is shown to be essential. The data are analysed 

in a model established for cross-relaxation in polymer hydrogels [4,5] . 

Employing Goldman-Shen experiments, the cross-relaxation effect can 

be extracted, and diffusion data re-interpreted. 

As a result, the stacked layers are found to contain micropores. The 

presence of these pores is confirmed by AFM images, and their size 

dependence on relative humidity is quantified from the NMR data. 

References: 

[1] A. Jin, A. Toutianoush, B. Tieke, Appl. Surf. Sci. 246 (2005) 444. 

[2] X. Liu, M. L. Bruening, Chem. Mater. 16 (2004) 351. 

[3] F. Vaca Chávez, M. Schönhoff, J. Chem. Phys. 126 (2007) 104705. 

[4] L.J.C. Peschier et al, J. Magn. Reson. B 110 (1996) 150. 

[5] D. Topgaard, O. Söderman, Langmuir 17 (2001) 2694. 

Td12 

Shear effects on surfactant mesophases as seen by NMR 

Medronho, Bruno 1 ; Ar, Gonul 2 ; Miguel, Maria C. 1 ; Olsson, Ulf 3 ; Schmidt, 

Claudia 2 

1 University of Coimbra, Department of Chemistry, Coimbra, Portugal; 

2 University of Paderborn, Department of Chemistry, Paderborn, 

Germany; 3 Lund University, Physical Chemistry, Lund, Sweden 

Shear can have pronounced effects on the orientation and structure 

of surfactant mesophases, leading to rheological phenomena such as 

shear-thinning or shear-thickening. In order to better understand the 

rheological properties of surfactant systems, many researchers combine 

rheological measurements with structural investigations. The majority 

of such studies uses scattering methods and direct optical observations 

[1]. NMR investigations are not yet very common in this field [2-4], 

although NMR spectrometers are much more accessible than large 

scale scattering facilities. In this contribution we will illustrate the use 

of NMR, focusing on the shear-induced structural transitions of the 

lyotropic lamellar phase [1,5,6]. The lamellar phase under shear may 

consist of extended planar lamellae that can have different orientations 

or of close-packed multilamellar vesicles (MLVs). The stability regions 

of these structures as a function of shear rate and temperature can 

be readily obtained from the 2 H NMR line shapes of D 2 O-enriched 

samples and mapped in a shear diagram. An analysis of the line 

widths and of diffusion properties can provide further structural details, 

such as the MLV size. Finally, transient processes under shear, such 

as the formation of MLVs from initially planar lamellae or the reverse 

process, can be followed. From such time-dependent NMR experiments, 

performed on the nonionic surfactant system C 10 E 3 /D 2 O, a 

fundamental difference depending on the direction of the transformation 

process has been found: the transition from planar lamellae to MLVs is 

a continuous transformation, whereas the opposite transition occurs via 

a “two-phase” region [6]. 

[1] S. Koschorek, S. Fujii, and W. Richtering, Prog. Theor. Phys. Suppl. 

2008, 175, 154-165. 

[2] P. T. Callaghan, Rep. Prog. Phys. 1999, 652, 599-670. 

[3] P. T. Callaghan, Current Opinion in Coll. Interface. Sci. 2006, 11, 

13-18. 

[4] C. Schmidt, in “Modern Magnetic Resonance”, Vol. 3, 1495-1501, 

Springer, New York. 

[5] O. Diat, D. Roux, and F. Nallet, J. Physique II 1993, 3, 1427-1452. 

[6] B. Medronho, S. Shafaei, R. Szopko, M. G. Miguel. U. Olsson, and C. 

Schmidt, Langmuir 2008, 24, 6480-6486. 


Td13 

Diffusion NMR studies of some ferrocenyl amidinium 

compounds: comparison of solution and solid state structure 

Stokes, Francesca A; Day, Iain J; Hitchcock, Peter B; Coles, Martyn P 

University of Sussex, Department of Chemistry and Biochemistry, 

Brighton, United Kingdom 

The ferrocenyl substituent, CpFe(η-C 5 H 4 )-, has found wide application 

as a reporter group used to probe molecular interactions. 1,2 The 

combination of the ferrocenyl reporter and a nitrogen-containing 

cationic amidinium group has recently been developed for the 

recognition of organic and inorganic oxo-anions. This recognition can 

be observed using a number of physical techniques including infrared 

spectroscopy and cyclic voltametry. 

X-ray crystallographic studies have shown a variety of hydrogen 

bonding motifs adopted by these compounds in the solid state, 

depending mainly on the relative bulk of the oxo-anion. Monomers, 

dimers and polymeric species in which the hydrogen bonding 

interaction is propagated in 1- or 2-dimensions have been observed. 

We have employed convection-compensated pulsed gradient stimulated 

echo (PGStE) NMR methods 3 to probe the molecular association state 

of a number of these compounds in the solution state. 4 Comparison of 

the hydrodynamic radii obtained from the diffusion NMR experiments 

in chloroform-d1 solution with computational results obtained using 

DFT shows that in solution state the likely aggregation states consist 

of smaller molecular fragments, with little evidence for the formation of 

long range order or the presence of larger multimeric species observed 

in the solid state. 

References 

(1) P.D. Beer, P.A. Gale, Z. Chen, Adv. Phys. Org. Chem., 1998, 31:1-90 

(2) P.D. Beer, P.A. Gale, Z. Chen, Coord. Chem. Rev., 1999, 185-186:3- 

36 

(3) A. Jerschow, N. Muller, J. Magn. Reson., 1997, 125:372-375 

(4) A. Macchioni, G. Ciancaleoni, C. Zuccaccia, D. Zuccaccia, Chem. 

Soc. Rev., 2008, 37:479-389 

Td14 

Using q-space and the homogeneous length scale to obtain 

structural information 

Åslund, Ingrid; Lasic, Samo; Söderman, Olle; Topgaard, Daniel 

Lund University, Physical Chemistry, Lund, Sweden 

The q-space analysis for the pulsed-field-gradient spin-echo (PGSE) 

has been used to obtain structural information for almost two decades. 

However, the conventional protocol to analyze the data presumes that 

the short-gradient-pulse (SGP) approximation holds. For technical 

reasons this is very rarely true. 

We have developed a technique that takes advantage of the break 

down of the SGP-approximation to obtain structural information of 

the examined system. By keeping the q-values used constant and 

changing the gradient pulse duration in a set of PGSE experiments a 

homogeneous length scale, L, can be obtained [1]. This is done through 

the q-value at which the echo attenuations for the different gradient 

pulse lengths starts to deviate from each other. L (= 1/q for the start 

of the deviation) is the length scale at which the system goes from 

appearing inhomogeneous to homogeneous. 

The inhomogeneities can be caused by a lot of different things. It can be 

domains with different diffusion coefficients and/or concentrations but 

also more complex things like confinements or anisotropic systems. 

By some prior knowledge of the possible structure of the system 

examined, L can be used to obtain actual structural sizes of the system. 

Without prior knowledge one always gets out the length scale for 

the inhomogeneities in the system making it possible to qualitatively 

ABSTRACTS POSTER


compare the structure of similar samples. This second feature makes it 

good to combine with imaging sequences to identify inhomogeneities in 

the structural features of a sample. 

[1] Åslund et al, J. Phys. Chem. 112 (2008) 2782-2794 

Td15 

New pure shift and pure shift DOSY experiments 

Aguilar Malavia, Juan A. 1 ; Evans, Robert 1 ; Nilsson, Mathias 1 ; Morris, 

Gareth A. 1 ; Haiber, Stephan 2 

1 University of Manchester, School of Chemistry, Manchester, United 

Kingdom; 2 Givaudan, Dept. for Analytical Research, Naarden, 


The ability to resolve signals is fundamental to any form of 

spectroscopy, including NMR. Overlapping signals complicate spectral 

analysis, and in the case of DOSY can severely distort the diffusion 

dimension. In 1 H spectra, signal overlap can be greatly reduced by 

suppressing the multiplet structure caused by homonuclear coupling. 

In contrast with heteronuclear broadband decoupling, where highly 

efficient techniques are available, broadband homonuclear decoupling 

is much more problematic. Most of the published schemes are 

unsatisfactory either because the signals obtained are non-quantitative, 

or because the resolution is poor, or both. Probably the best option, the 

Zangger-Sterk experiment (1), went virtually unnoticed until recently (2- 

4). This experiment produces quantitative, phase-sensitive, broadband 

homodecoupled spectra, albeit at a high cost in S/N. 

We have recently speeded up the Zangger-Sterk experiment 

substantially, and have demonstrated its potential in DOSY (2). We 

now present some new families of pure shift 1D NMR and pure shift 

DOSY pulse sequences, which increase both the versatility of this class 

of experiment and the quantity and quality of chemical information it 

provides, and make clear the underlying kinship between seemingly 

disparate pure shift methods such as BIRD (5), anti-z-COSY (6) and 

phase-sensitive 2DJ spectroscopy (3). 

1 K. Zangger and H. Sterk, J. Magn. Reson. 1997.124: 486 

2 M. Nilsson and G.A. Morris, Chem. Commun., 2007. 933 

3 A.J. Pell and J. Keeler, J. Magn. Reson. 2007. 189:293 

4 N. Giraud, M. Joos, J. Courtieu and D. Merlet, Magn. Reson. Chem. 

2009. 47:300 

5 J.R. Garbow, D.P. Weitekamp and A. Pines, Chem. Phys. Lett. 1982. 

93:504 

6 A.J. Pell, R.A.E. Edden and J. Keeler, Magn. Reson. Chem. 2007. 

45:296 

Td16 

Diffusion NMR and trilinear analysis in the study of reaction 

kinetics 

Khajeh, Maryam; Botana, Adolfo; Nilsson, Mathias; Bernstein, Michael 

A; Morris, Gareth A 


Kingdom 

Time-resolved NMR spectroscopy can in principle allow every species 

involved in a chemical reaction to be monitored simultaneously, 

providing both real-time quantitation and information on chemical 

structure. However, it is not always simple to interpret the data obtained, 

because where signals overlap it is difficult to distinguish between the 

signals of different components. In principle, statistical methods such 

as PCA can be applied to separate the components. Unfortunately, this 

typically results in rotational ambiguity, where a wide range of candidate 

component spectra fit the experimental data equally well. Multi-linear 

126 

analysis (where the data vary independently in more than 2 dimensions) 

offers a way around this problem, allowing experimental data to be 

decomposed into physically realistic component spectra where such 

data can be obtained experimentally. 

One way to obtain trilinear NMR data for the course of a chemical 

reaction is to acquire successive DOSY [1] (Diffusion-Ordered 

SpectroscopY) datasets, in which pulsed field gradients are used to 

attenuate the signals of different species according to their diffusion 

coefficients, during the reaction. After Fourier transformation, each 

individual DOSY dataset records how the NMR spectrum varies with 

pulsed field gradient strength at a given time. Provided that each 

species has a different diffusion coefficient and a different timecourse, 

the dataset is trilinear and can be decomposed using the PARAFAC [2] 

(PARAllel FACtor Analysis) algorithm to yield the spectrum, concentration 

time course, and diffusional attenuation for each component of the 

reaction separately[3]. 

Examples will be shown for model systems in which spectra are 

extracted, and accurate kinetic data obtained, even where the spectra 

are almost completely overlapped. 

1. G.A. Morris, in Encyclopedia of Nuclear Magnetic Resonance, eds. D. 

M. Grant and R. K. Harris, John Wiley & Sons Ltd, Chichester, 2002, vol. 

9 : Advances in NMR, pp. 35-44 

2. R. Bro, Chem. Intell. Lab. Syst., 1997. 38: p. 149-171. 

3. M. Nilsson, M. Khajeh, A. Botana, M. A. Bernstein, and G.A. Morris, 

Chem. Comm. 2009. p. 1252-1254 

Td17 

Diffusion ordered NMR spectroscopy to monitor the hydrolysis 

of p-nitrophenyl phosphate promoted by a heptamolybdate 

cluster 

Van Lokeren, Luk 1 ; Cartuyvels, Els 2 ; Absillis, Gregory 2 ; Willem, Rudolph 1 ; 

Parac-Vogt, Tatjana 2 

1 Vrije Universiteit Brussel, High Resolution NMR Centre, Brussels, 

Belgium; 2 Katholieke Universiteit Leuven, Department of Chemistry, 

Leuven, Belgium 

The half-life of phosphoester bonds, linking all nucleotide monomers 

in the DNA chain, is estimated at 1.3×10 5 by neutral pH and 298K, 

meaning that almost one billion years are required for complete 

hydrolysis of the DNA backbone. This kinetic stability is an excellent 

advantage important for the preservation of the genome. However, it 

certainly is a disadvantage when it comes to the repair of damaged 

DNA or to the destruction of foreign DNA. Polyoxometalates (POMs) 

are extensively investigated as an important family of metal-oxygen 

clusters. Since such metal complexes accelerate the hydrolysis of 

phosphoester bonds up to four orders of magnitude, they are widely 

used in catalysis, medicine and material science. The mechanism of 

this hydrolysis is generally described as an interplay of several factors. 

Among other criteria, the complex has to have an overall positive 

charge. However, the cleavage of phosphoesters by highly negative 

POM clusters was recently likewise reported. In this study the hydrolysis 

of a DNA model system, p-nitrophenyl phosphate (NPP), promoted by 

the polyoxomolybdate cluster [Mo 7 O 24 ] 6- was investigated. During the 

reaction, there is absolutely no free NPP in solution and two transient 

complexes are generated, giving finally rise to free phosphate and 

p-nitrophenol. 1D 1 H and 31 P NMR allowed to characterize one of the 

complexes as [(NPP) 2 Mo 5 O 21 ] 4- but did enable us to assign the second 

complex to either (NPP)Mo 7 O 25 (H 2 O) or [(NPP)Mo 6 O 18 (H 2 O) 3 ] 4- . 

Diffusion Ordered NMR Spectroscopy (DOSY NMR) made it possible to 

identify effectively these catalytically active transient clusters through 

their diffusive behaviour. Available size and shape information of the 

complexes allowed to estimate the diffusion coefficient of the species in 

solution. Comparing estimated and experimental diffusion coefficients 

confirmed the characterization of the first complex as [(NPP) 2 Mo 5 O 21 ] 4- 


and to identify the second complex as [(NPP) 2 Mo 12 O 36 (H 2 O) 6 ] 4- , a dimer 

of (NPP)Mo 6 O 18 (H 2 O) 3 . 

In conclusion, the combination of kinetic and diffusion based NMR 

results made it possible to propose a reaction mechanism for the 

hydrolysis of NPP catalyzed by a heptamolybdate. 

Td18 

The DOSY Toolbox: a new tool for processing PFGNMR 

diffusion data 

Nilsson, Mathias; Gareth A, Morris 


Kingdom 

The importance of high resolution PFGNMR data for mixture analysis 

is steadily increasing, but there is no single best way to process such 

data. The commonest family of processing methods is known as 

DOSY (diffusion-ordered spectroscopy), and therefore it has become 

customary to refer to these data as DOSY data. The three major NMR 

manufacturers each offer different limited implementations of DOSY 

processing in their current software. The DOSY Toolbox is a free 

programme that allows users of all three instrument families access to 

the same wide range of processing schemes. 

The DOSY Toolbox has a graphical user interface for easy access to 

the main processing schemes, and a command line mode for more 

advanced options. It is written in MATLAB, but is also available as 

free-standing compiled version that does not require any MATLAB 

installation. 

Basic features include: 

Import of Varian, Bruker and JEOL data, Weighting, phasing, baseline 

correction and referencing, Reference deconvolution 

DOSY data processing includes: 

DOSY (mono-, bi-, and multiexponential), Correction for non-uniform 

field gradients, DECRA, MCR, SCORE 

The DOSY Toolbox is released under the GNU public licence (GPL); a 

copy can be downloaded from http://personalpages.manchester.ac.uk/ 

staff/mathias.nilsson/ or requested by email from mathias.nilsson@ 

manchester.ac.uk. 

Td19 

Spectral analysis, time-dependent diffusion and diffusant 

relaxation rate in porous systems 

Nordin, Matias 1 ; Nyden, Magnus 1 ; Nilsson Jacobi, Martin 2 

1 Chalmers University of Technology / Applied Surface Chemistry, 

Chemical and Biological Engineering, Gothenburg, Sweden; 2 Chalmers 

University of Technology / Complex systems group, Energy and 

Environment, Gothenburg, Sweden 

An eigenfunction expansion of the Laplace operator in porous systems 

connects diffusion parameters to standard spectral analysis. Three 

parameters; tortuosity, surface-to-pore volume ratio and relaxation rate 

are derived from the spectrum of the Laplace operator and connected 

to the parameters in the Padé approximation, an expression often used 

to describe the time-dependent diffusion constant in porous systems. 

The Padé length is identified for systems with large pore to connector 

volume ratio. The results are compared with simulations. 


Td20 

Resolution of Isomer Spectra by Matrix-Assisted DOSY 

Evans, Robert 1 ; Haiber, Stephan 2 ; Nilsson, Mathias 3 ; Gareth A., Morris 3 

1 University of Manchester, School of Chemistry, Oxford Road, 

Manchester, United Kingdom; 2 Givaudan, Dept Analyt Res, Huizerstr, 

Naarden, Netherlands; 3 University of Manchester, School of Chemistry, 

Oxford Road, Manchester, United Kingdom 

Diffusion-ordered NMR spectroscopy (DOSY) is a very useful tool 

in the analysis of mixtures, separating the NMR signals of different 

components according to diffusion coefficient, but relies on different 

species having sufficiently different hydrodynamic radii. The great 

majority of DOSY experiments to date have used solutions of mixtures 

in simple solvents, but there is considerable scope for manipulating 

relative diffusion coefficients by the use of modified solvents. There is 

a close analogy here between DOSY and chromatography. Much of the 

richness of chromatography as an analytical tool stems from the subtle 

degree of control allowed by varying the nature of the stationary phase; 

the introduction of co-solvents and -solutes, to create a complex matrix 

in which differential diffusion between solutes is measured, gives DOSY 

a comparable flexibility. 

One simple and vary effective manipulation of the diffusion matrix 

in DOSY is to use a surfactant co-solute above its critical micelle 

concentration. This allows species of very similar size, such as isomers, 

to be distinguished from one another by virtue of their differing degrees 

of interaction with the micellar structures formed. 

Excellent resolution of the proton spectra of isomers can be obtained 

by using common surfactants such as SDS in aqueous solution. 

Good diffusion resolution is retained over a much wider range of 

concentrations of solute and surfactant than would be expected on 

the basis of a simple model of solute binding to micelles. The same 

principle may be exploited in nonaqueous solvents by using reversed 

micelles, such as those of AOT in chloroform. Here good resolution 

is once again readily obtainable, with added flexibility afforded by the 

ability to vary the water content of the reversed micelles. 

Td21 

Diffusion by PFG-NMR: The spy reporting from inside industrial 

samples 

Voelkel, Ruediger 

BASF SE, Polymer Physics, Ludwigshafen, Germany 

Diffusion measurements by pulsed field gradient (PFG) NMR are shown, 

discussing the “why” and “how” of various applications of PFG-NMR 

in research in chemical industry. It is our strategy to extend the use 

of PFG-NMR data beyond the aspects of transport in order to gain 

insight into the physico-chemical processes that occur within a sample. 

Diffusion may thus be the probe to report a change in the effective size 

of the molecule or particle, either by physical processes (adsorption, 

association, aggregation, micellization, exchange) or by chemistry 

(degradation of a polymer, reaction, complexation). 

- Transport: Diffusion measurements of 1% of ethylbenzene inside 

a polystyrene melt (200°C) are shown [1] as an example in which 

diffusion data are needed to model a technical process, i.e. the 

degassing of the residual monomer from a polystyrene melt by strand 

degassing. 

- Size of nano particles: The diffusion coefficient D of nano particles 

in a colloidal sample can be used to calculate their size by the 

Stokes-Einstein relation. The advantage of PFG-NMR over competing 

techniques as well as its limitations are discussed and examples of 

inorganic and organic particles shown. 

- Emulsions: The micellization of amphiphilic molecules and the 

exchange between micelles and monomeric emulsifier is accessible. 

PFG outperforms particularly, if the size distribution of droplets in suspo- 

ABSTRACTS POSTER


emulsions [2] is to be determined. 

- Spectroscopy: The PFG-NMR experiment may serve as a 2filter2 

to either purge spectra or group the signals belonging to the same 

chemical entity. The presentation is based on examples taken from 

various fields of industrial PFG-NMR. 

[1] V.A.Harmandaris, N.P.Adhikari, N.F.A.van der Vegt, K.Kremer, 

B.A.Mann, R.Voelkel, H.Weiss, CheeChin Liew (2007) Macromolecules 

40, 7026-7035 

[2] Jianqin Zhuang, R.Voelkel (2005) 5th annual surface and colloid 

symposium “Amphiphilic Polymers”, Lund; dto, Diffusion Fundamentals, 

3, 37.1-37.2 

Td22 

Memory effects in confined fluids via diffusion measurement 

Naumov, Sergej 1 ; Valiullin, Rustem 1 ; Kärger, Jörg 1 ; Monson, Peter A. 2 

1 Leipzig University, Dept. of Interface Physics, Leipzig, Germany; 

2 University of Massachusetts, Dept. of Chemical Engineering, Amherst, 

MA, United States 

For bulk fluids it is always possible to establish the state of equilibrium 

coexistence between vapour and liquid phase by placing samples of 

these phases in contact. On the opposite, the fluids under mesoporous 

confined may reside in several quasi-equilibrium coexistence states 

of different densities for very long time. The transition between these 

metastable states occurs via changes in the driving force (pressure or 

chemical potential, temperature of the bulk phase) or via the thermally 

induced fluctuations of the fluid. The latter process is generally 

accompanied by very large barriers in free energy and may result in the 

occurrence of the so called adsorption hysteresis. 

The hysteresis is a well-known feature of adsorption/desorption 

isotherms for light gases, such as nitrogen, at cryogenic temperatures, 

and, depending on the pore structure of the adsorbent, may involve 

the existence of a very large number of completely reproducible but 

non-equilibrium states that are accessible via scanning sorption 

experiments, i.e. various cycles of incomplete filling/draining 

procedures. Traditional adsorption measurements give the relationship 

between the confined fluid density and the bulk chemical potential (via 

the bulk pressure) but do not provide detailed information about the 

density distribution within the system. 

In this work, self-diffusion measurements by pulsed field gradient 

nuclear magnetic resonance (PFG NMR) are used to probe the state 

of a fluid, confined within the internal pore space of a model porous 

material, namely Vycor porous glass. 

We provide direct experimental evidence that states within hysteresis 

loops that have the same average fluid density may have different 

average self-diffusivities, as a direct reflection of differences in the 

density distributions between the states. Thus, molecular diffusivity 

is shown to be an excellent probe of the history-dependent states 

of the confined fluid. The self-diffusivities reflect different arrested 

spatial distributions of the confined fluid that accompany the very slow 

equilibration of the system in this region. 

128 

Td23 

Rapid acquisition of diffusion-diffraction q-space spectra from 

red blood cells: simulations and new applications 

Larkin, Timothy 1 ; Pages, Guilhem 1 ; Torres, Allan 2 ; Kuchel, Philip 1 

1 University of Sydney, School of Molecular and Microbial Biosciences, 

Sydney, Australia; 2 University of Western Sydney, Nanoscale 

Organisation and Dynamics Group, Campbelltown, Australia 

The rapid acquisition of q-space spectra from 1 H 2 O undergoing 

restricted diffusion in suspensions of red blood cells (RBCs) is made 

possible by using a recently implemented pulse sequence [1]. To 

decrease the time of each experiment, the phase cycling of the radiofrequency 

(RF) pulses is reduced to two transients with unbalanced 

pairs of bipolar gradient pulses [2]. The q-space spectra obtained show 

a shift in the position of the first diffraction minimum when compared to 

the classical pulsed field gradient stimulated echo (PGSTE) experiment. 

Monte-Carlo random walk simulations of the diffusion of water in a 

lattice of RBCs were used to investigate the effect of the additional 

delay introduced by the bipolar gradient pulses on the form of the 

q-space plots. 

The time saving enabled using the rapid-acquisition pulse sequence has 

opened new avenues of investigation. RBCs of normal discocyte shape 

align with an external magnetic field, and the angular dependence of 

q-space spectra from suspensions of RBCs was examined using a linear 

combination of gradients applied along the y and z-axes. The resulting 

q-space plots showed the gradual disappearance of the first diffraction 

minimum as the angle at which the gradients were applied was 

changed from 0° (along the z-axis) to ~35°, beyond which the q-space 

plots showed no diffraction features. These experimental results were 

confirmed by Monte-Carlo random walk diffusion simulations on RBCs 

with varying alignment with respect to B 0 . 

The general principle of this experiment and its analysis have 

implications for the interpretation of diffusion-weighted and q-space 

MRI images from in-vivo systems with non-symmetrical diffusionrestricting 

cellular and tissue compartments. 

1 Pages, G., Szekely, D. and Kuchel, P. W. (2008) Erythrocyte-shape 

evolution recorded with fast-measurement NMR diffusion-diffraction. J. 

Magn. Reson. Imag. 28, 1409-1416 

2 Pelta, M. D., Morris, G. A., Stchedroff, M. J. and Hammond, S. J. 

(2002) A one-shot sequence for high-resolution diffusion-ordered 

spectroscopy. Magn. Reson. Chem. 40, S147-S152 

Td24 

A combined NMR Diffusometry, TEM and brownian simulation 

investigation of the microstructure in alginate gels 

Bernin, Diana 1 ; Goudappel, Gert-Jan 2 ; Ruijven, Marjolein-van 2 ; Ström, 

Anna 2 ; Hazekamp, Johan 2 ; Hermansson, Anne-Marie 3 ; Nydén, Magnus 1 

1 Chalmers, DEPARTMENT OF CHEMICAL AND BIOLOGICAL 

ENGINEERING, Göteborg, Sweden; 2 Unilever, Research, Vlaardingen, 

Netherlands; 3 SIK, Göteborg, Sweden 

The combination of NMR diffusometry and TEM (Transmission Electron 

Microscopy), has been shown to be a valuable tool to study the 

intrinsic dependence between microstructure and diffusion of probe 

molecules in polymer gels. Dendrimers of different sizes (generations) 

are known to be good diffusion probes in polymer gels due to their 

well-defined size and spherical geometry. Alginate gels with different 

polymer concentrations and CaCO 3 concentrations have been prepared. 

In addition to the CaCO 3 dependence the microstructure was altered 

by additions of small amounts of methanol. The TEM images of the 

different gels show a variation in microstructure. According to TEM 

images methanol has only a small effect of the microstructure of the gel 

but the diffusometry experiments reveal significant changes in diffusion 


ehaviour of dendrimers. The results indicate that there are structures 

in gels not noted by the TEM method, although it provides nanometer 

resolutions information. The obtained diffusion coefficients have been 

used to characterise the microstructure of the polymer gels. 

Td25 

Diffusion analysis of structured and unstructured proteins 

Delsuc, Marc-André 1 ; Tanty, Matthieu 1 ; Cheynier, Véronique 2 ; Sarni- 

Manchado, Pascale 2 ; Paté, Franck 2 

1 CNRS, IGBMC - Biomolecular NMR, Strasbourg, France; 2 INRA, SPO, 

Equipe Polyphénols-Interactions, Montpellier, France 

Translational diffusion measurement by NMR (DOSY) has been used 

to study the hydrodynamic behaviour of structured and intrinsically 

unstructured proteins. 

In a recent work(1), it has been shown that diffusion NMR spectroscopy 

is a efficient technique to measure the spacial extension of polymer 

and protein chains in solution in terms of fractal dimension. The fractal 

dimension is a notion which measure the way a fractal object fills-up 

the 3D space. We have thus shown that structured proteins have a 

mean fractal dimension of 2.56 (very close to the highest possible value 

of 3.0); while denatured proteins display a fractal dimension of 1.71, 

thus behaving like polymers in a good solvent. The purpose of this work 

is to show that hydrodynamics and fractal dimension are pertinent 

descriptors of intrinsically unstructured proteins. 

This approach has recently been applied to several protein known to 

be intrinsically unstructured. First, the polyproline polymer has been 

studied. Measurements on a series of peptides have permitted to 

determine a fractal dimension compatible with a rather rigid, extended 

polyproline II secondary structure. 

The proline rich salivary protein IB5 is known to bind polyphenol. 

Unbound, it presents a rather extended structure, and its oligomerisation 

upon binding was monitored by DOSY(2). 

The AB domain of nuclear receptors are known to be disordered. ABdomain 

of RARα presents a proline rich domain which binds to SH3 

domains. This domain was compared to polyproline, and the modulation 

of its properties upon phosphorylation was monitored both by DOSY and 

chemical shift. 

1)S Augeì, PO Schmit, CA Crutchfield, MT Islam, DJ Harris, E Durand, 

M Clemancey, AA Quoineaud, JM Lancelin, Y Prigent, F Taulelle, MA 

Delsuc “NMR Measure of Translational Diffusion and Fractal Dimension. 

Application to Molecular Mass Measurement.” J.Phys.Chem B (2009) 

113 (7) 1914 

2)C Pascal, F Paté, MA Delsuc, V Cheynier, “Study of the interactions 

between a proline rich protein and a flavan-3-ol by NMR: residual 

structures of the natively unfolded protein are anchorage points for the 

ligands.” Biopolymer (2009) in press 

Td26 

Diffusion tensor imaging of water dynamics in roots of Zea Mais 

‘Helix’ 

Menzel, Marion I. 1 ; Spindler, Natascha 2 ; Blümich, Bernhard 3 ; Pohlmeier, 

Andreas 2 ; Vereecken, Harry 2 ; Schurr, Ulrich 1 

1 Forschungszentrum Jülich, Phytosphere Institute, Jülich, Germany; 

2 Forschungszentrum Jülich, Agrosphere Institute, Jülich, Germany; 

3 RWTH Aachen, Macromolecular Chemistry, Aachen, Germany 

Water uptake and transport are essential for plant nutrition. High 

resolution magnetic resonance imaging (MRI) allows unique options 

for non-invasive investigations of water flow and growth processes 

in root-soil-systems. Especially for the determination of slow water 

movements diffusion tensor magnetic resonance imaging (DTI) is 

convenient. This measures the effective diffusion coefficients of water 

in Cartesian directions in each pixel of an MR image. For the first time 

this technique is applied to roots to identify water mobility within roots. 

Anisotropic diffusion is expected because of the non-spherical shape 

of the different cell types (xylem, phloem elements) in roots. Based on 

DTI measurements also the preferential direction of water motion within 

roots relative to the laboratory coordinate system can be obtained which 

might help for a closed reconstruction of the root skeleton, necessary 

in future model calculations on root water uptake. For practical and 

visualization purposes it is convenient to reduce the diffusion tensor 

to a single parameter: the fractional anisotropy in each pixel, which is 

calculated from the eigenvalues of the diffusion tensor in each pixel. 

For the experiments we used three weeks old plants of Zea Mais ‘Helix’ 

cultivated in quartz sand (medium grain size: 0.36 mm) covered by a 

layer of coarse sand for the seed. All NMR images were taken using a 7 

T vertical magnet system (Varian), equipped with a micro imaging system 

(maximal gradient strength: 300 mT/m) and a 38 mm inner diameter 

birdcage resonator. Anatomical images and DTI were acquired using a 

conventional spin-echo multi-slice sequence with additional application 

of diffusion encoding gradients in different spatial directions and varying 

gradient strengths. The next step of the investigations is the detection 

of water motion in soil towards the roots. Also for this DTI might be 

convenient, since flow velocities are expected to be too small for direct 

flow imaging methods. First results on water diffusivity measured by 

means of DTI in medium sand are also included in the presentation. 


Td27 

Phase transitions in surfactant model systems: A spatially 

resolved rheo-NMR approach 

Medronho, Bruno 1 ; Brown, Jennifer R. 2 ; Galvosas, Petrik 2 

1 Department of Chemistry, University of Coimbra, Coimbra, Portugal; 

2 MacDiarmid Institute for Advanced Materials and Nanotechnology, 

Victoria University of Wellington, Wellington, New Zealand 

The lyotropic lamellar phase of surfactant solutions may exhibit 

interesting structural transformations under shear flow. Most interesting 

is the formation of monodisperse, close-packed, multilamellar vesicles 

(MLVs), which are referred to as onions [1]. 

Such transformations have been reported recently for the nonionic 

surfactant model system of C 10 E 3 in D 2 O. These results were obtained 

in a rheo-NMR cylindrical Couette with deuterium NMR spectroscopy 

which uses the quadrupole interaction of D 2 O as a probe for the state of 

orientation and order parameter. The transformation from planar layers 

to onions was found to be a slow continuous/homogeneous and strain 

controlled process. On the other hand, the reverse transition from onions 

to planar lamellae was found to be a much faster discontinuous process 

[2]. 

In this contribution we extend previous research on the same surfactant 

system by the combination of NMR spectroscopy and diffusometry with 

magnetic resonance imaging (MRI). Chemical shift imaging revealed the 

discontinuous nature of the transition from onions to planar lamellae, 

confirming the spatial coexistence of both phases within the gap of 

the couette geometry as predicted in [2]. On the other hand, NMR 

diffusometry revealed a strain dependence of the diffusion and the 

spatial direction of the diffusive process strongly suggests that a multilamellar 

cylindrical or undulated intermediate structure exists during 

the transition from the lamellar to the onion phase. Moreover, in our 

experiments, the orientation of this intermediate structure is aligned 

along the flow direction as reported by Zipfel et al. [3]. 

[1] O. Diat, D. Roux and F. Nallet, J. Physique IV 3 (C8), 193-204 (1993). 

[2] B. Medronho, S. Shafaei, R. Szopko, M. G. Miguel, U. Olsson and C. 

Schmidt, Langmuir 24, 6480-6486 (2008). 

[3] J. Zipfel, F. Nettesheim, P. Lindner, T. D. Le, U. Olsson and W. 

Richtering, Europhys. Lett. 53, 335-341 (2001). 

ABSTRACTS POSTER

AUTHOR INDEX 

Aachmann, Finn L. Bi45*, Bi46* 

AB, Eiso Mi28* 

Abergel, Daniel Co11* 

Absillis, Gregory Td17 

Adams, Alina Im11, Sn13* 

Adams, Stephen R. So26 

Aebi, Markus Bi47 

Aeby, Nicolas Re26 

Aganov, Albert Sp14 

Agarwal, Sunil Im10 

Agarwal, Vipin Pl07 

Aguilar Malavia, Juan A. Td15* 

Ahuja, Puneet En03, Re24 

Airoldi, Cristina Mi26* 

Akasaka, Kazuyuki Bi31 

Akke, Mikael Re11 

Al-Bawab, Abeer Sm19 

Alcocer, Marcos Mi18 

Alexenskii, Alexander Sp02 

Al-Hashimi, Hashim M. Pl01* 

Allix, Mathieu Sn41 

Amero, Carlos So04* 

Amoureux, Jean-Paul Sn22, Sn29 

Andersson, K. Kristoffer Ep13 

André, João P. Mi24 

Ansermet, J.-Ph. En18 

Ansermet, Jean-Philippe En03 

Antti, Henrik Co13 

Antzutkin, Oleg N. Sn35* 

Anwar, Muhammad Sabieh En17* 

Ar, Gonul Td12 

Araya, Yonathan Im04 

Arcon, Denis Ep20, Sp04 

Ardenkjær-Larsen, Jan H. En26 

Arnqvist, Anna Bi22 

Arrowsmith, Cheryl Co03 

Arseniev, Alexander Bi04, Bi15, Bi24, Bi25, Bi44, Bi57, 

Bi73, So43 

Åslund, Ingrid Td03, Td14* 

Aspers, Ruud Sm28* 

Atkinson, Andrew So30 

Atkinson, Kevin En04 

Auzanneau, France-Isabelle Sm17 

Axmann, Marco Mi11 

Babailov, Sergey Pa12* 

Bade, Steffen Bi50 

Bains, Ravi Ep11 

Baishya, Bikash Re26 

Baldus, Johanna Sn35 

Banci, Lucia So22 

Bardiaux, Benjamin Bi67, Co18 

Bargon, Joachim En15, En16 

Barths, Daniela Bi50 

Báthori, Mária Sm26 

Bauer, Birgitta Sn33 

Bauer, Christian Ep11 

Baum, Marina Bi69 

Baumann, Herbert En26 

Bayrhuber, Monika Mi32 

Beck, Irene En21 

Beck, Johannes G. Sm23* 

Becker, Stefan Bi02, Bi33, Ep22, Mi32, Pa10, 

Re02, Sn11 

Bednarek, El¿bieta So29 

Behera, Babita Sn10 

Belov, Alexander Pa13 

Benjamin, Nathan Pa15 

Bennati, Marina En23, En24, Ep01*, Ep22 

Berger, Robert Sm18 

Berglund, Lars A. Im15 

Bermel, Wolfgang So25 

Bernadó, Pau Mi02 

Bernin, Diana Im16, Td24* 

Bernstein, Michael A Td16 

Berthault, Patrick En12 

Bertini, Ivano Pa03, So19, Sn28, So22 

Bhargava, Satish Im10 

Bhatt, Shuchi Im10 

Bhaumik, Anusarka Sn28 

Bibow, Stefan So40* 

Biernat, Jacek Sn11, So40 

Biesemans, Monique Sn53 

Billeter, Martin Co18, Co22, So25 

Birkan, Burak So35 

Bjerring, M. Sn04 

Björk, Heikki So27 

Björlenius, Berndt Me02 

Blackledge, Martin Re01*, Re03, So31 

Blanchard, Laurence So31 

Blechta, Vratislav Sm14 

Blindow, Silke Bi50 

Blommers, Marcel Bi04* 

Blümich, Bernhard Sn03*, Td26 

Blümler, Peter Re21 

Bobnar, Matej Sn33* 

Bocharov, Eduard Bi15*, Bi24, Bi25, Bi57, So43 

Bocharova, Olga Bi04, Bi15, Bi24 

Bochtler, Matthias Bi27 

Bocian, Wojciech So29 

Böckmann, Anja Sn01*, Sn15, Sn28 

Bodenhausen, Geoffrey Co11, En03, Im02, Re24, Re26, 

So03 

Bohle, Anne Sn14* 

Böhme, Ute Mi22, Re18 

Boisbouvier, Jerome So04 

Bonnal, Sophie Mi10 

Bontems, François Bi13, Mi31 

Bonvin, Alexandre Co18 

Boos, Winfried Bi49 

Bordignon, Bordignon Mi29 

Bordignon, Enrica Ep03, Pa14 

Boros, Sándor So38* 

Botana Alcalde, Adolfo Td10*, Td16 

Böttcher, Benjamin Sm11*, Sm18 

Bouchard, Louis-S. En17 

Boucher, Wayne Co18 

Bousset, Luc Sn01, Sn15 

Bovee-Geurts, Petra H. M. Sn02 

Brasun, Justyna Bi29 

Bren, Kara L. Ep13 

Broadhurst, Bill Co10* 

Brose, Nils Bi33 

Brotin, Thierry Re27 

Brown, Jennifer R. Td27 

Brown, Richard C. D. Sn02 

Brown, Steven P. Sn35 

Brückner, Ann-Kathrin Bi02 

Brunklaus, Gunther Sn14, Sn17 

Brunner, Konrad Co18 

Brus, Jiri Sn50 

Brusova, Hana Sn50 

Brutscher, Bernhard So04 

Bryndal, Iwona Sn47 

130 *Presenting author

Buchinger, Edith Bi46 

Buchner, Johannes So41 

Büchner, Bernd Sp15 

Bukhtiyarov, Valery En21 

Bunge, Andreas Sn18 

Burkhardt, Ulrich Sn33 

Burschowsky, Daniel Bi14* 

Burt, Scott R. En17 

Buschhorn, Sabine Bi01, Mi30 

Butterwick, Joel A. Ep21 

Cadars, Sylvian Sn20 

Calandrini, Vania Co11 

Calle, Luis Pablo Bi54* 

Cametti, Massimo So36 

Campbell, Iain Bi61 

Cañada, Francisco Javier Bi54 

Cañana, J. Mi24 

Cantù, Laura Mi26 

Capek, Peter Bi56 

Carella, Michela Bi69 

Carepo, MS So19 

Caretti, Ignacio Ep16 

Carlomagno, Teresa Bi33 

Carlos Dias, Luiz Sm21 

Carlsson, Jonas Co26* 

Carravetta, Marina Pl03, Sp03 

Carrigan, John Me03 

Cartuyvels, Els Td17 

Carvalho, Eugenia Sm24 

Casanova, Federico Sn03 

Casanueva, Felipe F. Bi28 

Castro, M. Margarida Sm24* 

Cavazzini, Davide Bi12 

Cebrat, Marek Bi29 

Cecowski, Saša Sn51 

Cefaro, Chiara So22 

Cekan, P. Ep25 

Cevec, Mirko Bi32* 

Chakraborty, Swagata So20* 

Chan Huot, Monique Bi34* 

Chao, Yu Bi23 

Chapelle, Anne So30 

Charpentier, Thibault Sn22, Sn30 

Chary, Kandala V So13 

Chattah, Ana Karina Sn42, Sn43 

Chaturvedi, Shubhra Me10 

Chechik, Victor Re22 

Cherepanov, Alexey Sn12* 

Cheung, Nicole Co10 

Chevelkov, Veniamin Pl07 

Cheynier, Véronique Td25 

Chi, Chunhwa Bi37 

Chia, Winlong Sn19 

Chizhik, Vladimir Co14, So11 

Cho, Hyun-soo Bi37 

Choi, Yun-Seok Bi72 

Chupin, Vladimir Bi04 

Cindric, Marina Sm27 

Ciobanu, Luisa Im12* 

Ciofi-Baffoni, Simone So22 

Claridge, Timothy D. W. Bi35 

Clayden, Nigel Sn39 

Clerico, Eugenia M. So21 

Coles, Martyn P Td13 

Collins, Tony So32 

Colombo, Laura Mi26 

Comment, Arnaud En03, En18 

Concistre, Maria Sn02* 

Conlon, Michael So23 

Corsini, Lorenzo Mi23 

Coskun, Ünal Bi01, Mi30 

Costa Pessoa, João Sm24 

Costanzo, Stéphanie So31 

Cottrell, Stephen Sn39* 

Coudevylle, Nicolas Bi33, Pa02 

Coville, Neil Ep15 

Cowley, Michael En04 

Craven, C Jeremy So44 

Cristol, Sylvain Sn22, Sn30 

Csikós, Orsolya Bi63 

Cuda, Francesco Sp03 

Cuffini, Silvia Sn42, Sn43 

Cukier, Cyprian D. Bi43* 

Cumpstey, Ian Sm12 

Cuny, Jerome Sn27* 

Curro, Nick J. Sp15 

Dafforn, Tim Bi01 

Dahlberg, Carina Im03* 

Dalibor, Paar Sp15* 

Damberg, Peter So26 

Dames, Sonja A. Bi36* 

Danieli, Ernesto Sn03 

Danielsson, Jens Bi42 

Danquigny, Alain Sp03 

Dasari, Muralidhar Pl07 

Daughdrill, Gary W. So31 

d’Auvergne, Edward Pa02*, Sm21 

Davis, Benjamin G. Bi35 

Day, Iain J Td13* 

De Grip, Willem J. Sn02 

de la Hoz, Antonio Fr01 

De Paëpe, Gaël Sn28 

Dedek, Matt Sm23 

Del Favero, Elena Mi26 

Delevoye, Laurent Sn22, Sn27, Sn30* 

Delgado, Teresa Sm24 

D’Elia, Valerio Sm03 

Delsuc, Marc-André Td25* 

Demers, Jean-Philippe Sn11 

Dempwolff, Felix Bi49 

Denisov, Gleb So28 

Denysenkov, V. En02, En22, Ep25* 

Deschamps, Michael Sn20 

Desvaux, Hervé En12* 

Dhulesia, Anne Co11 

Díaz Hernández, María Dolores Bi28* 

Diaz-Moreno, Irene Bi43 

Díaz-Ortíz, Ángel Fr01 

Diehl, Anne Sn16 

Diehl, Annette Bi67 

Diehl, Carl Re11* 

Diercks, Tammo En27* 

Diez Berart, Sergio Sn45 

Dikiy, Alexander Bi45 

Dilger, Herbert Ep14 

Döbber, Meike Ep03 

Doedens, Lucas Sm23 

Doglia, Silvia Maria Mi26 

Dolinšek, Janez Ep23, Sn33, Sp12 

Dollmann, Björn En11 

Domján, Attila Sn44* 

Donets, Alexey So11* 


AUTHOR INDEX

AUTHOR INDEX 

Dong, Ronald Sn36* 

Doreleijers, Jurgen Co18 

Doronin, Serge Co12 

Doroteya, Staykova Co22 

Dorr, Brent Pa02 

Drahus, M. D. En28 

Drašar, Pavel So36 

Drettwan, Diana Mi13* 

Drosten, Christian Mi11 

Dubosclard, Virginie Bi13 

Duckett, Simon En04* 

Duerst, Matthias Bi69 

Dufourc, Erick J. Sn31 

Dupree, Ray Sn35 

Dürr, Ulrich H. N. So42* 

Dutasta, Jean-Pierre Re27 

Duus, Jens Ø. En26, Mi33 

Dvinskikh, Sergey Im15* 

E. Kövér, Katalin So38 

E. Unveren, Elif So35 

Eddy, Matthew T. Sn28 

Eden, Mattias Sn34* 

Efremov, Roman Bi15, Bi25, Bi57 

Egbe, D.A.M. Sp14 

Eichel, R.-A. En28 

El Shawish, Samir Ep20 

Eleouet, Jean-François Bi13 

Eliav, Uzi Fr02 

Emsley, Lyndon Pa03, Pa04, Re03 

Endeward, Burkhard En02, En22, Ep21* 

Engelhard, Martin Ep03 

Engelke, Frank En23, En24 

Ensign, Amy A. Ep13 

Erdelyi, Mate Bi68 

Erdem, E. En28* 

Eremina, Rushana Ep24* 

Ernst, Matthias Sn25 

Erünal, E. En28 

Ervasti, Mia So27 

Espy, Michelle Im04 

Esteban-Martín, Santi Bi17 

Esteban-Martín, Santiago Co17* 

Eva, Selstam Mi18 

Evans, Robert Td15, Td20* 

Falk, Kirsten En14 

Fallis, Ian A. Ep16 

Faneca, Henrique Sm24 

Fares, Christophe Co03, Pa02 

Farkas, Daniel Mi27* 

Farkas, Viktor Bi63 

Fassbender, Birgit Sn17* 

Fayon, Franck Sn20 

Fazlizhanov, Ilshat Ep24 

Federwisch, Guido Mi13 

Fedorova, Anna Co12 

Feintuch, Akiva Ep19 

Fel’dman, Edward Co12* 

Feliz, Miguel En20 

Felli, Isabella Pa03 

Fenwick, Bryn Co17 

Fenwick, Robert Bi03*, Re02 

Feuerbacher, Michael Sn33 

Figaroa, Francis Mi28 

Filipek, Renata Bi27 

Filippov, Andrei Sn35 

Finka, Rachael Bi01 

Finotello, Daniele Sn45 

Fleischmann, Matthias Sm03* 

Florian, Pierre Sn20, Sn41 

Flynn, Mark Im04 

Fogh, Rasmus Co18 

Forler, Nina Sn30 

Förlin, Lars Me02 

Foster, Mark Mi19* 

Frank, Benedikt Pa02 

Fransson, Iris G. Mi03 

Frantsuzov, Ilya Re15* 

Franzoni, Lorella Bi12 

Franzoni, María Belén Sn49* 

Fratila, Raluca Fr01 

Fredriksson, Jonas Co22*, So25 

Freund, Christian En14 

Friberg, Anders Mi23* 

Fritzinger, Bernd Mi17 

Fruh, Virginie Mi28 

Frydman, Lucio Fr02 

Fujara, Franz Re12 

Fujito, Teruaki Sn46 

Furó, István Im03, Im15, Td01* 

Gabel, Frank En19 

Gabellieri, Cristina En20 

Gábor, Tóth Bi63 

Gafurov, Marat En02, En22* 

Gairí, Marga Mi02 

Gajda, Jaroslaw Sn32, Sn47* 

Gallo, Angelo So22* 

Gallopin, Matthieu Re10 

Galvosas, Petrik Pl09*, Td27* 

Ganesan, A. Pl03 

Gang, Ga-Ae Bi51 

Ganin, Alexey Y. Sp04 

Gans, Pierre So04 

Gardiennet, Carole Sn01, Sn15 

Gareth A, Morris Td18, Td20 

Garro Linck, Yamila Sn42*, Sn43* 

Gärtner, Tobias Sm15 

Gautier, Regis Sn27 

Ge, Min Sp03 

Geahlen, Robert Bi11 

Geissler, Erik Sn44 

Genheden, Samuel Re11 

Geraldes, Carlos F.G.C. Mi24*, Sm24 

Gergely, András Sm26 

Gerum, Christian Bi20 

Gescheidt, Georg En25 

Ghalebani, Leila Bi53*, Re19 

Gierasch, Lila M. So21 

Giesler, Reiner So12 

Gil, Sergi So17*, So18* 

Gille, Peter Sn33 

Giller, Karin Mi32 

Gillis, Pierre Pa16, Re23 

Giraud, Pierre Mi31* 

Gladden, Lynn Im01* 

Gladyshev, Vadim N. Bi45 

Glaser, S.J. Sn04 

Glaubitz, Clemens Sn26, Sn37 

Gnezdilov, Oleg Mi20 

Goerlach, Matthias Bi59, Bi69 

Goldfarb, Daniella Ep04, Ep19, Pl05* 

Gollnick, Paul Mi19 


Gomez, John Im04 

Gomez, M. Victoria Fr01* 

Gómez-Caravaca, Ana María So33 

Gonçalves, Gisela Sm24 

Goncharuk, Marina Bi15, Bi57, So43 

Gossuin, Yves Pa16*, Re23 

Goudappel, Gert-Jan Td24 

Govan, Norman Sm22 

Graf, Robert Sn43 

Grafe, Hans-Joachim Sp15 

Granwehr, Josef En13 

Gräslund, Astrid Bi42, Bi53, So26 

Graumann, Peter Bi49 

Green, Gary En04 

Griesinger, Christian Bi02, Bi03, Bi33, Bi68, Co19, En23, 

En24, Ep22, Mi32, Pa02, Pa10, 

Re02, Sm21, So40 

Griesser, Markus En25* 

Griffin, Julian Me01* 

Griffin, Robert G. Sn28 

Grin, Yuri Sn33 

Gröbner, Gerhard Bi22, Sn31, So12 

Gronwald, Wolfram Sm03 

Grosse, Frank Bi59 

Grossel, Martin Sp03 

Grzesiek, Stephan Bi36, Co02 

Gschwind, Ruth M. Mi13, Sm03, Sm15 

Guenther, Sebastian En14 

Guigas, Bruno Co18 

Guittet, Eric Re10 

Günther, Ulrich En10, Me03 

Guo, Jing So28* 

Gutmanas, Aleksandras Co03* 

Haag, M. En18 

Haarmann, Frank Sn33, Sp01* 

Habeck, Michael Mi32 

Habenstein, Birgit Sn01, Sn15 

Hackl, Thomas Mi03* 

Haglund, Ellinor Bi52 

Hagn, Franz So41 

Haiber, Stephan Td15, Td20 

Halle, Bertil Pl10*, Re04 

Hammerath, Franziska Sp15 

Hansen, Poul Erik Bi30 

Hanson, Christina Im04 

Hansson, Örjan Mi27 

Hanykova, Lenka Mi16 

Harbitz, Espen Ep13 

Hass, Mathias Pa01 

Hassani Nejad, Sayed Karim So10 

Hassinen, Tommi Bi21 

Hautle, Patrick En03, En18 

Hazekamp, Johan Td24 

Heetebrij, Rob Mi28 

Heggen, Marc Sn33 

Hegyi, Orsolya Bi63 

Heiles, Sven Sm04 

Heinicke, Friedericke Bi19 

Heller, Jeannine Ep02 

Hellmich, Ute Andrea Sn37 

Henrick, Kim Co18 

Henriksson, Marielle Im15 

Herlach, Dierk Mi20 

Hermansson, Anne-Marie Td24 

Herrmann, Andreas Sn18 

Herrmann, Torsten Bi14, Co04* 

Hewage, Chandralal Bi74, So16, So23 

Heydenreich, Katja Bi23* 

Hiet, Julien Sn20 

Hilger, Daniel Bi48 

Hiller, Sebastian Co24 

Hilty, Christian Bi02 

Hinderberger, Dariush En11*, Ep02*, Ep11*, Ep12, Mi12* 

Hinderhofer, Markus Bi49 

Hirschinger, Jerome Sn42 

Hitchcock, Peter B Td13 

Hoepner, Sabine En14 

Höfer, Peter En23, En24 

Holden, Ian Sm22* 

Hollander, Johan Mi28 

Hollingworth, David Bi43 

Holterhues, Julia Ep03 

Hore, P. J. Bi26 

Horsewill, A. J. Re15 

Hosur, Ramakrishna V Bi38, So20 

Houben, Klaartje So31 

Hovav, Yonatan Ep19 

Hrabal, Richard Bi62 

Hrenar, Tomica Sm27 

Hruby, Victor J. Sm23 

Hu, Bingwen Sn29 

Huang, Jie-rong Co02* 

Huber, Gaspard En12 

Huber, Matthias Fr03 

Hung, Ivan Sn35 

Hunter, C Niel So44 

Huster, Daniel Sn18* 

Huttunen, Eine Bi16 

Huvonen, D. Sp03 

Hyz, Karolina So29* 

Ibraghimov, Ilghiz Co24 

Igor, Tkach En23 

Ilc, Gregor Bi29 

Ilg, Karin Bi47 

Inaba, Kenji Bi60 

Inukai, Munehiro Im14*, Sn52* 

Invernizzi, Chiara Mi29 

Ivanov, Konstantin So37 

Ivanshin, Vladimir Sp13* 

Iwai, Hideo Bi46 

Iyudin, Vasiliy Ep18* 

Jackson, Trudy A Sm17 

Jacquet, Eric Bi64 

Jaeger, Martin Sm28 

Jakes, P. En28 

James, Jonathan Bi01 

Janin, Sami En03, En18* 

Janssen, Hans WG Sn21 

Jansson, Mats So12 

Jaravine, Victor Co18, Co24 

Jaremko, Lukasz Bi27*, Bi29 

Jaremko, Mariusz Bi27, Bi29* 

Jarvet, Jüri Bi42 

Jeganathan, Sadasivam So40 

Jeglic, Peter Sp04*, Sp12 

Jegliè, Peter Sn33 

Jehle, Stefan Bi40, Bi67, Sn16 

Jelen, Andreja Ep23 

Jensen, Malene R. So31* 

Jensen, Pernille R. En26, Mi33 

Jeon, Young Ho Bi72* 


AUTHOR INDEX

AUTHOR INDEX 

Jerschow, Alexej Re17 

Jeschke, Gunnar Bi48*, Pa14* 

Jeziorna, Agata Sn32 

Jiang, Pengju Bi61 

Jimenez, Lucia Sn17 

Jiménez-Barbero, Jesús Bi28, Bi54, Mi24 

Johannessen, Ole G. Sn02 

Johansson, Conny M Mi25* 

Johansson, Pär Bi22 

John, Michael Sm16* 

Jokisaari, Jukka Re20, Sn48* 

Jones, John Sm24 

Jones, Jonathan A. Bi35 

Jovin, Thomas Ep22 

Jung, Heinrich Bi48 

Jur ek, Ond ej So36* 

Jurczak, Janusz So39 

Kajino, Tsutomu Bi60 

Kalbitzer, Hans Robert Co18 

Kalisiak, Jaroslaw So39 

Kamiya, Yukiko Bi60 

Kärger, Jörg Td22 

Karlsson, B. Göran Bi10 

Karlsson, Magnus En26, Mi33* 

Karyagina, Irina Ep22* 

Kashida, Shoji Sp10 

Kato, Koichi Bi60, Bi68 

Katrakili, Nitsa So22 

Kau i , Ven eslav Sn51 

Kauèiè, Venèeslav Pa11 

Kauppinen, Reijo Sn10 

Kaur, Ravinder Ep13 

Kawahara, Seiichi Sn38 

Kawêcki, Robert So29 

Kazimierczuk, Krzysztof Bi41, Co23* 

Kazmierski, Slawomir So39* 

Keartland, Jonathan Ep15*, Sp11* 

Kehlet, C. Sn04 

Keizers, Peter Pa01 

Kelly, Geoff Bi43 

Kemper, Sebastian Bi35* 

Kennedy, Gordon Sn23* 

Kentgens, Arno PM Sn21 

Kervern, Gwendal Pa04 

Kesselring, Eric Im13 

Kessler, Horst Sm01*, Sm23, So41 

KGS, K.G.Srinivasa Bi55 

Khajeh, Maryam Td16* 

Khaneja, N. Sn04, So01* 

Khantimerov, Sergei Mi20 

Khramtsov, Valery Im13* 

Khushu, Subash Me10 

Kieffer, Bruno Re13, So30 

Kilian, Wolfgang En14 

Kilpeläinen, Ilkka Bi61 

Kim, Eunice EunKyeong Bi72 

Kim, Hwa-Young Bi45 

Kim, Jae-Hong Bi72 

Kim, Ji Sun Bi71 

Kim, Sangyun Bi37, So15 

Kim, Yongae Bi51, Bi71* 

King, Glenn F. Bi58 

Király, Péter Sm20 

Kiraz, K. En28 

Kitahara, Ryo Bi31 

Kizilsavas, Gönül Mi12 

Klanjsek, Martin Ep20, Sn33 

Klapper, Markus Sn17 

Klare, Johann Ep03 

Kleber Z. Andrade, Carlos Sm21 

Kleckner, Ian Mi19 

Kleinmaier, Roland Mi13 

Kleshyov, Andrey En11 

Klevit, Rachel Bi67, Sn16 

Klinkhammer, Karl W. Ep12 

Kneller, Gerald Co11 

Knight, Michael Pl03 

Knowles, Timothy J Bi01 

Ko, Sunggeon Bi37*, So15 

Kobayashi, Masakazu Mi28 

Kobera, Libor Sn50 

Koch, Achim En15, En16 

Köck, Matthias Co19, Sm02* 

Köckenberger, Walter En13 

Koeppe, Benjamin So24*, So37 

Kolehmainen, Erkki So36 

Kolehmaninen, Erkki Sn10 

Kolodzik, Adrian Fr04 

Komatsu, Koichi Sp03 

Kondrsenko, A.A. Sm13 

Köneke, Stephanie Sn25 

Konkin, Alexander Sp14* 

Konovalov, A.A. Ep10* 

Konter, J.A. En18 

Konter, James En03 

Koptyug, Igor Co15, En17, En21 

Korhonen, Samuli-Petrus Bi21 

Kornievskaya, Valeria Mi21 

Korukottu, Jegannath So40 

Koskela, Harri So27* 

Koskela, Outi Bi61* 

Kotsyubynskyy, Dmytro Re19* 

Kourilova, Hana Mi16 

Kousik, Chandra So13* 

Kovtunov, Kirill En17, En21 

Kowalewski, Jozef Re19, Re27 

Kozminski, Wiktor Co01* 

Kozerski, Lech So29 

Kozminski, Wiktor Bi41, Co23, Sm25 

Kraus, Robert Im04 

Krishnamohan, Poluri Maruthi So20 

Kruk, Danuta Re12* 

Kruppa, Alexandr Ep18, Mi21 

Kuchel, Philip So02, Td02, Td23 

Kuhn, Lars T. Bi26* 

Kühne, Ronald Bi67 

Kukol, A. Sn35 

Kukovitsky, Eugene Mi20 

Kumar, Dinesh Bi38 

Kunjuzwa, Nikiwe Ep15 

Kurfurst, Milan Sm14 

Kurimoto, Eiji Bi60 

Kurzbach, Dennis Ep12*, Mi12 

Kwak, Sooyoung So15 

Laage, Ségolène Pa03* 

Laatikainen, Reino Bi21 

Labuta, Jan Mi16 

Lagleder, Stephan So41* 

Lahtinen, Manu Sn10 

Lakomek, Nils Bi03 

Lakomek, Nils-Alexander Re02 

Lamazhapova, Darima Co18 


Lamosa, Pedro So32 

Landström, Jens Mi15* 

Lang, Guillaume Sp15 

Lang, Jan Bi62 

Lange, Adam Sn11* 

Lantto, Perttu Co16* 

Larkin, Timothy Td02, Td23* 

Larsson, Göran Co13, Mi18 

Larsson, Joakim Co24, Me02 

Lasic, Samo Td14 

László, Krisztina Sn44 

Laue, Ernest Co18 

Lavrentovich, Oleg Sn45 

Le Bihan, Denis Im12 

Le Polles, Laurent Sn27 

Lebedev, Andrey Pa13 

Lee, Donghan Bi02*, Re02 

Lee, Inhwan So15 

Lee, Jae-Jin Bi72 

Lee, Kong-Joo Bi72 

Lee, Soo Hyung Ep17 

Lee, Weontae Bi37, So15 

Leffler, Hakon Re11 

Léger, Patrick Sn25 

Leggett, James En13* 

Lehtivarjo, Juuso Bi21* 

Lelli, Moreno Sn28 

Lemak, Alexander Co03 

Lenoir, Marc Bi01, Mi30* 

Leonov, Andrei Pa10 

Leppert, Joerg Bi69 

Lerche, Mathilde H. En01*, En26, Mi33 

Lesage, Anne Pa03 

Leshina, Tatyana Mi21* 

Levitt, Malcolm Pl03*, Sn02, Sp03 

Levstein, Patricia R. Sn49 

Lewandowski, Józef R. Re03*, Sn28* 

Li, Lin Ep03 

Li, Yuzhuo Sm19 

Liebscher, Jürgen Sn18 

Liebscher, Markus Bi23 

Liktor-Busa, Erika Sm26 

Limbach, Hans-Heinrich Bi34, So24, So28, So37 

Lin, Yu-Pin Bi01 

Lind, Jesper Bi52*, Bi66, So26 

Lindfors, Hanna Pa01 

Lindorff-Larsen, Kresten Re16 

Linser, Rasmus Pl07 

Litvinova, Tatyana Sp13 

Lo, Andy Y H Sn34 

Loch, Caroline Mi28 

Locshin, Alexander Mi20 

Lodi, Alessia Me03 

Longhi, Sonia So31 

Lopes, Philippe Co11 

Lopez, Jakob J. Sn26* 

López, Paloma Bi54 

Loquet, Antoine Sn01, Sn15*, Sn28 

Lounila, Juhani Re20* 

Louzao, Iria Sm18 

Low, Check-Fong Bi58 

Lowry, David F. So31 

Lu, Yi-Zhi Sn19 

Luchinat, Claudio Pl04*, Sn28 

Lücke, Christian Bi12*, Bi23 

Ludwig, Christian En10*, Me03 

Lukzen, Nikita Co15* 

Lyubenova, Sevdalina En22 

Lyukmanova, Ekaterina Bi44*, Bi73 

Lyukmanova, Katya Bi04 

Maas, Werner Sn28 

Mackereth, Cameron En19 

MacKinnon, Roderick Ep21 

Madani, Fatemeh So26* 

Maddinelli, Giuseppe Re14* 

Madl, Tobias Bi50, En19* 

Maeno, Aya Bi60 

Maestre-Martinez, Mitcheell Bi33* 

Magnelind, Per Im04 

Maguire, Mahon Co10 

Mainz, Andi Bi40* 

Mäler, Lena Bi52, Bi65, Bi66 

Mali, Gregor Pa11*, Sn51 

Maliniak, Arnold Sm18 

Malm, Linus Co13* 

Malthouse, J.Paul G Bi74 

Mamone, Salvatore Pl03, Sp03* 

Mandelkow, Eckhard Sn11, So40 

Manoj, Manoj Bi55* 

Manolikas, Theofanis Fr03 

Manthey, Katarina Sp15 

Manzo, Rubén H. Sn42, Sn43 

Manzoni, Claudia Mi26 

Marassi, Francesca M. Fr04 

Marchand, Philipp Bi64* 

Margraf, D. Ep25 

Mari, Silvia Mi29 

Marie, Sabrina Pl03 

Marin-Montesinos, Ildefonso En10, Me03* 

Marion, Denis J.Y. En12 

Marion, Dominique So31 

Marko, A. Ep25 

Markovic, Stefan Bi67*, Sn16 

Marquardsen, Thorsten En23, En24 

Martell, Swetlana Ep03 

Martinelli, Manuele So22 

Martín-Pastor, Manuel Bi28 

Martins, José A. Mi24 

Martins, Jose C. Mi17*, Re13 

Marx, Andreas Sm10* 

Maskaly, Karlene Im04 

Massiot, Dominique Sn20*, Sn40, Sn41 

Matlashov, Andrei Im04 

Matulova, Maria Bi56* 

Matzapetakis, Manolis So32* 

Matzen, Guy Sn41 

Mauri, Francesco Sn22, Sn30 

Maury, Olivier Pa04 

Mayzel, Maxim Bi15, Bi57* 

McDermott, Ann Mi04 

McElroy, Craig Mi19 

McKenzie, Iain Ep14*, Sn39 

McLean, Neville Sn02 

Medronho, Bruno Td12, Td27 

Medycki, Wojciech Re12 

Mehl, Georg Sn48 

Mehrotra, Gopesh Im10 

Meier, Beat Sn15 

Meier, Beat H. Bi47, Fr03, Sn01, Sn25 

Meier, Sebastian En26*, Mi33 

Meins, Thomas Mi32 

Meirovitch, Eva Re19 

Melacini, Giuseppe Bi18* 


AUTHOR INDEX

AUTHOR INDEX 

Melki, Ronald Sn01, Sn15 

Mennes, Nadine Ep03 

Menzel, Marion I. Td26 

Meyer, Bernd Mi03, Mi11 

Meyer, Franc Sm16 

Meyer, Helge Bi50* 

Miguel, Maria C. Td12 

Miñana, Bela Mi10 

Mineev, Konstantin Bi15, Bi25*, Bi57 

Minying, Cai Sm23 

Mishkovsky, Mor Fr02 

Misiak, Maria Sm25* 

Mitschang, Lorenz En14* 

Mobli, Mehdi Bi58* 

Mohoriè, Aleš Td04 

Möller, Heiko M. Bi49* 

Monson, Peter A. Td22 

Montagne, Lionel Sn22, Sn30 

Monti, Gustavo A. Sn42, Sn43 

Montouillout, Valérie Sn20, Sn41* 

Moreno, Andrés Fr01 

Morris, Gareth Td10, Td15, Td16 

Moudrakovski, Igor Sn24* 

Mountford, Philip So34 

Moura, I So19 

Moura, JJG So19 

Mourao, Andre Mi10*, Mi23 

Moza, Sahil Me10* 

Mueller, Norbert Re17* 

Mugnaini, Veronica En20* 

Mukovskii, Yakov Ep24 

Mukrasch, Marco D. So40 

Münnemann, Kerstin En11, En15, En16 

Murata, Yasujiro Sp03 

Murphy, Damien M. Ep16 

Musco, Giovanna Mi29* 

Nadezhdin, Kirill Bi15, Bi24* 

Nagarajan, Vijayasarathi Ep19* 

Nagel, U. Sp03 

Nakano, Michiko Bi60 

Nakasako, Masayoshi Bi60 

Narayanan, Raghav L. So42 

Natalello, Antonino Mi26 

Nath, Pulak Im04 

Naumann, Christoph So02* 

Naumov, Sergej Td22* 

Nausner, Martin Re17 

Navarini, Luciano Bi56 

Navarro-Vázquez, Armando Bi70, Co21* 

Navon, Gil Fr02* 

Neese, Frank Pl08* 

Nekrasova, Oksana Bi44 

Neshchadin, Dmytro En25 

Neumeier, Maria Sm15* 

Newman, Shaun Im04 

Nicotra, Francesco Mi26 

Niederweis, Michael Fr04 

Nielsen, A.B. Sn04 

Nielsen, J.T. Sn04 

Nielsen, N.C. Sn04 

Nikonov, Georgii So34 

Nilges, Michael Co18, En19 

Nilsson, Markus Td03 

Nilsson, Mathias Td10, Td15, Td16, Td18*, Td20 

Nilsson, Ulf Re11 

Nilsson Jacobi, Martin Td19 

Nolis, Pau So17, So18 

Nonappa, Sn10* 

Nordin, Matias Td19* 

Nordstierna, Lars Im16, Re28* 

Novak, Predrag Sm27* 

Novak Tušar, Nataša Sn51 

Novikov, Valentin Pa13* 

Nowacka, Agnieszka Td03 

Nowicka, Katarzyna So39 

Nydén, Magnus Im16, Td19, Td24 

Obynochny, Anatoly Ep18 

Oddone, Anna Mi23 

Odeh, Fadwa Sm19* 

O’Harte, Finbarr So16 

Ohki, Shinobu Sn46 

Ohlenschlaeger, Oliver Bi59*, Bi69 

Olaru, Alexandra Maria Im11* 

Olejniczak, Sebastian Sn47 

Oliveberg, Mikael Bi52, Bi65 

Oliveira Rocha, Rafael Sm21 

Olivera, María Eugenia Sn42, Sn43 

Oliveros, Malena En20 

Olofsson, Annelie Bi22 

Olsson, Ulf Td12 

Opperer, Florian Sm23 

Orekhov, Vladislav Co18, Co24* 

Ory, Sandra Sn41 

Oschkinat, Hartmut Bi40, Bi67, Sn16 

Osipov, Vladimir Sp02 

Östlund, Åsa Im16* 

Otting, Gottfried Pl06* 

Ouazzani, Jamal Co11 

Ould M’hamed Ould Sidi, Abdelahi So30 

Ovchinnikova, Tatiana Bi44 

Overduin, Michael Bi01*, Mi30 

Overhand, Mark Pa01 

Owens, Tuba Im04 

Pages, Guilhem Td02*, Td23 

Pais, Tiago M So32 

Pallister, Peter Sn24 

Palmer, Erin Sm23 

Pandya, Maya Bi31 

Panek, Rafal En13 

Panich, Alexander Sp02*, Sp10* 

Paquin, Raphael Im02* 

Parac-Vogt, Tatjana Td17 

Paramonov, Alexander Bi73 

Parella, Teodor So17, So18, So33 

Park, Joon Kyu Bi72 

Park, Tae Joon Bi51, Bi71 

Passadouro, Marta Sm24 

Passerini, Cinzia Re14 

Paté, Franck Td25 

Patel, Mitul Bi35 

Paul, Jean-François Sn22, Sn30 

Pauleta, Sofia R. So19* 

Pavlova, Maria Co14 

Pavoni, Silvia Re14 

Pazos, Yolanda Bi28 

Pedersen, Anders Bi10*, Sn31 

Pellerin, Nadia Sn20 

Pelupessy, Philippe Co11, Im02, So03* 

Penkett, Chris Co18 

Peräkylä, Mikael Bi21 

Perczel, András Bi63 


Perez, Yolanda Mi02 

Pérez, Miriam So17, So18 

Perez-Linde, Angel En13 

Pérez-Trujillo, Miriam So33* 

Perlo, Juan Sn03 

Permi, Perttu Bi61 

Persson, Erik Re04* 

Persson, Per So12 

Persson Sunde, Erik Pl10 

Peter, Matthias Bi14 

Peters, Fabian Pa10* 

Peters, Mark Im04 

Peters, Thomas Mi15 

Peterson, I.V. Sm13* 

Petrova, Marina Co15 

Petrova, Svetlana Mi21 

Petrovskaya, Lada Bi04 

Petryakov, Sergey Im13 

Petzold, Katja Bi22*, Re16*, So14 

Pfefferle, Susanne Mi11 

Pickard, Chris J. Sn27 

Piculjan, Katarina Sm27 

Pierattelli, Roberta Pa03 

Pileio, Giuseppe Pl03, Sn02 

Pines, Alexander En17 

Pinoie, Vanja Sn53* 

Pintacuda, Guido Pa03, Pa04* 

Plavec, Janez Bi32 

Podkorytov, Ivan S Pa15 

Pohlmeier, Andreas Re21, Td26 

Polimeno, Antonino Re19 

Polyakov, Nicolai Mi21 

Polyansky, Anton Bi15 

Polyhach, Yevhen Bi48, Pa14 

Pons, Miquel En20, Mi02* 

Pospiech, Helmut Bi59 

Post, Carol Beth Bi11* 

Potapov, Alexey Ep04* 

Potocnik, Anton Ep20* 

Potrzebowski, Marek Sn32*, Sn47, So39 

Prandolini, M. J. En02* 

Prandolini, Mark En22 

Prassides, Kosmas Sp04 

Prchal, Jan Bi62 

Prisner, T. F. En02, Ep25 

Prisner, Thomas En22, Ep21 

Privalov, Alexei Re12 

Proudfoot, Andrew So44* 

Prudencio, Miguel Pa01 

Pruski, Marek Sn23 

Pustovalova, Yulia Bi15, Bi25, So43* 

Qvist, Johan Pl10 

Rademacher, Christoph Mi15 

Rajagopal, Ponni Bi67, Sn16 

Ralph, Adams En04 

Ramachandran, Ramadurai Bi69 

Ramos, Andres Bi43 

Rana, Poonam Me10 

Rangus, Mojca Sn51* 

Rao, D Krishna Pa15 

Ratajczyk, Tomasz Re25 

Rathgeb-Szabo, Klara Bi36 

Raya, Jesus Sn42 

Raza Khan, Ahmad Me10 

Rebek, Julius Jr. Bi42 

Rees, Nicholas So34* 

Reese, Marcel En23*, En24 

Rehbein, Kristina Bi67, Sn16 

Reif, Bernd Bi40, Pl07* 

Reinscheid, Uwe Co19*, Sm21 

Reiser, Oliver Sm03 

Ren, X. En28 

Renato, Longhi Mi29 

Rennella, Enrico So03 

Reynaud, Olivier Im12 

Richter, Barbara Bi03 

Richter, Christian Bi26 

Riedel, Dietmar Ep22 

Rieping, Wolfgang Co18 

Ripmeester, John Sn24 

Rissanen, Kari So36 

Ritter, Uwe Sp14 

Rizzardi, Paolo Mi29 

Roch, Alain Re23 

Rodriguez-Castaneda, Fernando Bi33, Pa02 

Roetzschke, Olaf En14 

Rohonczy, János Co20, Co25 

Rohonczy-Boksay, Erzsébet Co25 

Romañuk, Carolina Sn42, Sn43 

Rõõm, Toomas Sp03 

Roslund, Mattias Sm12 

Rosseinsky, Matthew J. Sp04 

Rossi, Gian Luigi Bi12 

Roth, Meike En15*, En16* 

Rothery, Emma Sm22 

Rovó, Petra Bi63* 

Rubailo, A.I. Sm13 

Rubcic, Mirta Sm27 

Rubtsov, Denis Me01 

Rudolf, Fabian Bi14 

Ruigrok, Rob W. H. So31 

Ruijven, Marjolein-van Td24 

Ruml, Tomas Bi62 

Rundqvist, Louise Mi18* 

Rybin, Vladimir So30 

Ryde, Ulf Re11 

Ryu, Kyoung-Seok Bi72 

Sachleben, Joseph Mi19, Pa03 

Sachse, Anna Sm16 

Saez, Natalie Bi58 

Sagdeev, Renad Co15 

Sakurai, Satoshi Sn38* 

Sal, Lena S. Bi45 

Salgado, Gilmar Co11 

Salgado, Jesús Bi17 

Salikhov, Kev Ep18 

Salmona, Mario Mi26 

Salvatella, Xavier Bi03, Co17, Re02 

Samadi-Maybodi, Abdolraouf So10* 

Samouilov, Alex Im13 

Samuelsson, Linda Co24, Me02* 

Sánchez-Pedregal, Víctor M. Bi70* 

Sandin, Henrik Im04* 

Sani, Marc-Antoine Sn31 

Santos, Helena So32 

Sariciftci, Serdar Sp14 

Sarkar, Riddhiman En03, Re24* 

Sarni-Manchado, Pascale Td25 

Sasakawa, Hiroaki Bi60 

Sattler, Michael Bi50, En19, Mi01*, Mi10, Mi23 

Sauerwein, Andrea Pl03 


AUTHOR INDEX

AUTHOR INDEX 

Saunders, Martin En10, Me03 

Savelov, Andrei Co15 

Savukov, Igor Im04 

Säwén, Elin Bi16*, Sm12 

Saxena, Krishna Bi26 

Schanda, Paul Fr03, So04 

Scharff, Peter Sp14 

Scheler, Ulrich Mi22*, Re18*, Sn40* 

Scheuermann, Robert Ep14, Mi20 

Schiemann, O. Ep25 

Schiene-Fischer, Cordelia Bi23 

Schlagnitweit, Judith Re17 

Schleucher, Jürgen Bi22, Me04*, Mi18, Re16, So14 

Schlottgayer, Anna Mi21 

Schlundt, Andreas En14 

Schmid, Markus Sm03 

Schmidt, Claudia Td12* 

Schmidt, Gesine Sm02 

Schmidt, Holger Pa10 

Schmidts, Volker Sm11, Sm18* 

Schneider, Annerose Bi59 

Scholz, Ingo Sn25* 

Schönhoff, Monika Td11 

Schraml, Jan Sm14* 

Schramm, Gabriele Bi50 

Schreiber, Laura En11 

Schuetz, Anne Sn01, Sn15 

Schuleit, Michael Im03 

Schulga, Alexey So43 

Schultz, Larry Im04 

Schurr, Ulrich Td26 

Schwalbe, Harald Bi19, Bi20, Bi26 

Sedenkova, Ivana Sn50 

Seema, S. Bi55 

Segawa, Takuya Re26* 

Segura-Carretero, Antonio So33 

Seiboth, Thomas Bi69* 

Sein, Julien Re03 

Serša, Igor Td04 

Serve, Olivier Bi60* 

Sezer, Deniz En02 

Shames, Alexander Sp02 

Sharma, Yogendra So13 

Shenderovich, Ilja G. Mi14* 

Shenkarev, Zakhar Bi04, Bi44, Bi73* 

Shet, Keerthi Im13 

Shimizu, Tadashi Sn46 

Shinagawa, Hideyuki Sn46* 

Shingarova, Lyudmila Bi73 

Sideris, Dionisa So22 

Siegal, Gregg Mi28 

Siemer, Ansgar Mi04* 

Sigurdsson, S. Th. Ep25 

Silvers, Robert Bi19, Bi20 

Simon, András Sm26 

Simon, Bernd En19 

Simons, Kai Bi01, Mi30 

Sindhuwinata, Nora Mi15 

Singh, Venus Bi38* 

Sinnaeve, Davy Re13* 

Sironi, Erika Mi26 

Sitkowski, Jerzy So29 

Sizun, Christina Bi13*, Bi64, Mi31 

Sjöström, Michael Co13 

Skjåk-Bræk, Gudmund Bi46 

Skrynnikov, Nikolai R. Pa15, Pl07 

Sleucher, Jürgen So12 

Smith, Corinne Bi01 

Smith, Mark E. Sn35 

Smock, Robert G. So21 

Smrecki, Vilko Re17, Sm27 

Smyth, D. M. En28 

Sobczuk, Adam So39 

Söderman, Olle Td14 

Sokolenko, W.A. Sm13 

Soltesova, Maria Re27 

Somer, M. En28 

Song, Jinsue Bi72 

Soós, Tibor Sm20 

Sourigues, Yannick Sn01, Sn15 

Spevacek, Jiri Mi16* 

Spiess, Hans W. En11, En15, En16, Sn14, Sn17, 

Sn43 

Spindler, Natascha Td26* 

Spink, Edward Bi74* 

Spitaleri, Andrea Mi29 

Srb, Pavel Bi62* 

Srivastava, Ajai Im10* 

Stapf, Siegfried Re21 

Starovoytova, Larisa Mi16 

Staykova, Doroteya Co18, So25* 

Steiner, Peter-René Co18 

Steinhoff, Heinz-Jürgen Ep03* 

Stepišnik, Janez Td04* 

Steuernagel, Stefan Pa03 

Stevens, Tim Co10 

Stevens, Timothy Co18* 

Stevensson, Baltzar Sm18 

Sticht, Jana En14 

Stingaciu, Laura-Roxana Re21* 

Stokes, Francesca A Td13 

Stoykov, Alexey Ep14 

Straasoe, L.A. Sn04 

Strandberg, Erik Bi17* 

Ström, Anna Td24 

Struck, Anna-Winona Mi11* 

Struppe, Jochem Sn28 

Subasinghage, Anusha Priyangika So23* 

Suggi-Liverani, Furio Bi56 

Sukhanov, Andrey Sp13 

Suleimanov, Nail Mi20* 

Sun, Han Sm21* 

Sun, Honglei Pa15 

Sun, Ziki Im13 

Sunnerhagen, Maria Co03 

Svensson, Mona Sm17* 

Svirskaya, N.M. Sm13 

Sychrovsky, Vladimir Sm14 

Sykora, Jan Sm14 

Szalay, Zsófia Co20*, Co25* 

Szczepanowski, Roman H. Bi27 

Szczygiel, Agnieszka Mi17 

Szekely, David Td02 

Szpryngiel, Scarlett Bi65* 

Szymanski, Slawomir Re25* 

Takabayashi, Yasuhiro Sp04 

Takacs, Zoltan Re27* 

Takács, Mária Sm26* 

Takeda, Kazuyuki Im14, Sn52 

Tallavaara, Pekka Sn48 

Tandon, Anupama Im10 

Tanty, Matthieu Td25 

Tarasov, V.F. Ep10 


Tárkányi, Gábor Sm20* 

Tavernier, Andreas En23, En24 

Tayler, Michael Pl03 

Teixeira, João C. Mi24 

Tengel, Tobias Mi18 

Teriete, Peter Fr04* 

Tervonen, Henri Re20 

Thiele, Christina Sm04*, Sm10, Sm11, Sm18, So14* 

Thureau, Aurelien Re10* 

Thureau, Pierre Pl03 

Timmermans, Leo Mi17 

Tkach, Elena Bi25 

Tkach, Igor En24 

Tkachev, Yaroslav Bi39* 

Tokatlidis, Kostas So22 

Tolstoy, Peter Bi34, So24, So28, So37* 

Tomaz, Ana Isabel Sm24 

Tomlinson, Jenny Bi30 

Topgaard, Daniel Td03*, Td14 

Torres, Allan Td02, Td23 

Tosner, Z. Sn04* 

Tóth, Gábor Sm26 

Tóth, Noémi Sm26 

Travé, Gilles So30 

Traversari, Catia Mi29 

Trebosc, Julien Sn27 

Trébosc, Julien Sn29* 

Trigo-Mouriño, Pablo Bi70 

Tripathi, Rajendra P Im10 

Tripsianes, Konstantinos Bi50 

Tsien, Roger Y. So26 

Tucker, Ian Ep14 

Turano, P. So19 

Turdu, Nursen So35* 

Türke, Maria-Teresa En23, En24* 

Tycko, Robert Pl11* 

Tzou, Der-Lii Sn19* 

Ubbink, Marcellus Pa01* 

Uhrín, Dušan Mi25 

Ullah, Saif Bi30 

Ulrich, Anne S Bi17 

Unnerståle, Sofia Bi66* 

Urbaitis, Al Im04 

Urbanova, Martina Sn50* 

Utsumi, Hiroaki Sn38 

V. Dvinskikh, Sergey Im03 

Vaara, Juha Co16 

Vakulya, Benedek Sm20 

Valcárcel, Juan Mi10 

Valiullin, Rustem Td22 

Valla, Svein Bi46 

van Beek, Jacco D. Sn25 

van den Brandt, B. En03, En18 

van der Drift, Anniek En13 

van der Klink, J.J. En03, En18 

van Dijk, Marc Co18 

Van Doorslaer, Sabine Ep16* 

van Dusschoten, Dagmar Re21 

van Eck, Ernst RH Sn21* 

Van Lokeren, Luk Td17* 

van Rossum, Barth-Jan Bi67, Sn16* 

vanHeijenoort, Carine Re10 

Vanninen, Paula So27 

Varga, Szilárd Sm20 

Varnai, Peter Re16 

Vasa, Suresh K Sn21 

Vasconcelos, Filipe Sn22*, Sn30 

Vasos, Paul R. En03*, Re24 

Veciana, Jaume En20 

Vega, Shimon Ep19 

Velders, Aldrik Fr01 

Vendruscolo, Michele Bi03, Re16 

Venneti, Kalyana So16* 

Venturi, Chiara Bi28 

Vereecken, Harry Re21, Td26 

Verel, Rene Fr03* 

Véron, Emmanuel Sn20, Sn41 

Vieth, Hans-Martin So37 

Vijayan, Vinesh Sn11 

Villinger, Saskia Mi32* 

Vincent, Andrea So12* 

Vinck, Evi Ep16 

Vinding, M.S. Sn04 

Vlach, Jiri Bi62 

Vlasie, Monica Pa01 

Voelkel, Ruediger Td21* 

Volegov, Petr Im04 

Volkov, Alexander Pa01 

Voloshin, Yan Pa13 

Volynsky, Pavel Bi15, Bi25, Bi57, So43 

Vonrhein, Clemens Mi32 

Vosegaard, T. Sn04 

Vovk, Mikhail Co14* 

Vranken, Wim Co18 

Vriend, Gert Co18 

Vrtnik, Stanislav Sn33, Sp12* 

Vuister, Geerten Co18 

Vul’, Alexander Sp02 

Vuong, Quoc Lam Pa16, Re23* 

Vyalikh, Anastasia Sn40 

Wagner, Gerhard Co24 

Wahlström, Anna Bi42*, Bi53 

Walker, Christina A. Bi49 

Wallgren, Marcus Sn31* 

Walter, Korvin Re02*, Bi02 

Wang, An Sm17 

Wang, Qiang Sn29 

Warsi, Muhammad Re22* 

Wasmer, Christian Sn01, Sn15 

Waterman, Claire Me01 

Webb, Andrew Im12 

Weichold, Oliver Im11 

Weihermüller, Lutz Re21 

Wencka, Magdalena Ep23* 

Wenckebach, W. Th. En18 

Wende, Christina Td11* 

Werner, Joern Pl03 

Werning, Laura Bi54 

Wider, Gerhard Bi14 

Widmalm, Göran Bi16, Mi15, Sm12*, Sm17 

Wiench, Jurek Sn23 

Willem, Rudolph Sn53, Td17 

Williamson, David En04 

Williamson, Mike Bi30*, Bi31*, So44 

Wilton, David Bi31 

Wimmer, Reinhard Bi46 

Wimmer, Zden k So36 

Wirmer-Bartoschek, Julia Bi19*, Bi20* 

Witte, David Bi49 

Wojciechowski, Marek Bi27 

Wollschlag, Sandra Johanna Sn37* 


AUTHOR INDEX

AUTHOR INDEX 

Wolter, Anja Sp15 

Wong, Alan Sn35 

Worrall, Jonathan Pa01 

Wrachtrup, Jörg Pl02* 

Wu, Bin Co03 

Xiang, Sheng-Qi So42 

Xu, Jiageng Mi19 

Xu, Xingfu Pa01 

Xue, Y. Pl07 

Xue, Yi Pa15* 

Y. Inana, Tulay So35 

Yagi, Hirokazu Bi68 

Yamaguchi, Takumi Bi68* 

Yamamoto, Masahiro Bi68 

Yamamoto, Sayoko Bi68 

Yang, Zhennai Bi16 

Yee, Adelinda Co03 

Yeom, Tae Ho Ep17* 

Ylänne, Jari Bi61 

Yu, Jeong-A Bi51* 

Yu, Jiho Bi37 

Yun, Ji Hye So15* 

Yurkovskaya, Alexandra So37 

Zalar, Bostjan Sn45 

Zamani, Sepideh Ep16 

Zandomeneghi, Giorgia Bi47* 

Zanier, Katia So30* 

Zawadzka, Anna Bi41*, Co23 

Zdunek, Janusz Re16 

Zenchuk, Alexandre Co12 

Zerbetto, Mirco Re19 

Zeth, Kornelius Mi32 

Zhang, L. X. En28 

Zhang, Xue Bi16 

Zhang, Yajie Bi11 

Zharikov, E.V. Ep10 

Zhivonitko, Vladimir En21* 

Zhukov, Igor Bi27, Bi29 

Zhuravleva, Anastasia So21* 

Zoppellaro, Giorgio Ep13* 

Zorko, Andrej Ep20 

Zotev, Vadim Im04 

Zupancic, Blaz Sn45* 

Zweckstetter, Markus Mi32, So40, So42 

Zweier, Jay Im13 



AUTHOR INDEX

5 

9 

10 

7 

6 

MAP OF GÖTEBORG 

1 

2 4 

1 Chalmers Conference Centre 2 Elite Park Avenue Hotel 

3 Quality Panorama Hotel 

4 Hotel Lorensberg 

9 Boats for Älvsborgs Fortress 5 Hotel Vasa 

10 City Hall Börsen 6 Hotel Poseidon 

7 Hotel Flora 

8 SGS Apartments (Hostel) 

3 

8

* Wiley Prize Winner 

19:00 19:00 

City Reception 

Conference Dinner 

Hospitality Suites - Bruker BioSpin 

Hospitality Suites - Varian Hospitality Suites - Jeol 

14:00 14:00 Poster session Biomolecules Small Molecules 

Poster session 

14:20 M Overduin H Kessler 

14:40 

14:40 

Runan Lee Köck 

Runan 

15:00 15:00 Frontiers I: V Gomez 

Fenwick Fleischmann 

Frontiers III: R Verel 

15:20 15:20 Frontiers II: G Navon Blommers Thiele 

Frontiers IV: P Teriete 

15:40 Runan 

15:40 

Coffee 

Coffee Coffee 

16:00 16:00 

Opening 

16:20 

16:20 

Relaxation Solid state Physics Molec. Interactions Computational 

M Levitt 

16:40 16:40 M Blackledge F Haarmann M Sattler W Kozminski 

The Andrew Prize 

17:00 

17:00 Walter Panich 

Pons Huang 

17:20 

17:20 

C Luchinat 

Lewandowski Mamone 

Hackl Gutmanas 

The Russel Varian Prize 

17:40 

17:40 

Persson* Jeglic 

Siemer Herrmann 

Groupment Ampere General Assembly 

18:00 18:00 

Excursion 

Closing 

R Tycko 

12:00 12:00 Amero Duckett Potapov Sandin Tosner Pintacuda Schleucher Stepisnik 

Vendors 

12:20 12:20 

12:40 12:40 

Lunch 

Lunch Lunch 

Lunch 

Users 

13:00 13:00 

13:20 13:20 

Meetings 

13:40 13:40 

11:00 11:00 N Khaneja M Lerche M Bennati L Gladden A Böckmann M Ubbink J Griffin I Furo 

Registration Open 

11:20 11:20 Naumann Prandolini Hinderberger Paquin Concistre D'Auvergne Samuelsson Pages 

11:40 11.00 - 16.30 11:40 Pelupessy Vasos Steinhoff Dahlberg* Blümich Laage* Marin-Montesinos Topgaard 

10:40 10:40 Solution NMR Enhanced NMR EPR Imaging Solid State NMR Paramagnetic Metabolomics Transport&Diffusion 

Runan Palmstedt Runan Palmstedt Runan Palmstedt Runan Palmstedt 

10:00 

10:00 

Coffee Coffee Coffee Coffee 

09:15 09:15 

J Wrachtrup 

G Otting 

F Neese 

B Halle 

08:30 08:30 

H al-Hashimi 

D Goldfarb 

B Reif 

P Galvosas 

Runan Runan Runan Runan 

Sunday 5 

Monday 6 Tuesday 7 Wednesday 8 

Thursday 9 

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