RESEARCH REPORT 2003 I 2004 - FMP Berlin

FORSCHUNGSINSTITUT
FÜR MOLEKULARE PHARMAKOLOGIE
IM FORSCHUNGSVERBUND BERLIN E.V.
RESEARCH REPORT
2003 I 2004

FORSCHUNGSINSTITUT FÜR
MOLEKULARE PHARMAKOLOGIE
Campus Berlin-Buch
Robert-Rössle-Str. 10
D-13125 Berlin
fon: +49-(0)30-94793-100
fax: +49-(0)30-94793-109
e-mail: maul@fmp-berlin.de
Outstation:
Department of Molecular Genetics
FEM Steglitz
Krahmerstr. 6-10
D-12207 Berlin
fon: +49-(0)30-8437-1910
fax: +49-(0)30-8437-1922
The Forschungsinstitut für Molekulare
Pharmakologie (FMP) evolved out of the
Institut für Wirkstoffforschung (IWF) of the
Academy of Sciences of the GDR. Together
with seven other research institutions, the
institute is administratively organized
within the Forschungsverbund Berlin e.V.
The FMP is a member of the Leibniz
Association.
Das Forschungsinstitut für Molekulare
Pharmakologie ist aus dem Institut für
Wirkstoffforschung (IWF) der Akademie
der Wissenschaften der DDR hervorgegangen.
Es ist mit sieben weiteren
Forschungsinstituten im Forschungsverbund
Berlin e.V. zusammengeschlossen.
Das FMP ist ein Mitglied der
Leibniz-Gemeinschaft.
RESEARCH REPORT 2003 / 2004
Editorial Board:
Walter Rosenthal, Michael Bienert,
Ivan Horak, Hartmut Oschkinat
Compilation & Editing:
Björn Maul
Cover Design and Layout:
Hoch3 GmbH, Berlin
Photos:
Bernhard Schurian, Thomas Oberländer,
Björn Maul, Dana Hausbeck
FMP
Print:
Druckhaus Berlin-Mitte
This Research Report is also available at
www.fmp-berlin.de
Cover: The G-Protein coupled receptor
GPR 109: binding site with ligand
(nicotinic acid)
FORSCHUNGSINSTITUT
FÜR MOLEKULARE PHARMAKOLOGIE
IM FORSCHUNGSVERBUND BERLIN E.V.

SCIENTIFIC ADVISORY BOARD
WISSENSCHAFTLICHER BEIRAT
Professor Dr. Rudolf Balling
GBF - Gesellschaft für Biotechnologische Forschung mbH
Mascheroder Weg 1
38124 Braunschweig
Professor Dr. Annette G. Beck-Sickinger
Institut für Biochemie der Universität Leipzig
Brüderstr. 34
04103 Leipzig
Professor Dr. Matthias Bräutigam
Schering AG
Müllerstr. 170-178
13342 Berlin
Professor Dr. Christian Griesinger
Max-Planck-Institut für Biophysikalische Chemie
Am Faßberg 11
37077 Göttingen
Professor Dr. Reinhard Jahn (Vorsitz)
Max-Planck-Institut für Biophysikalische Chemie
Am Faßberg 11
37077 Göttingen
Professor Dr. Hans-Georg Joost
Deutsches Institut für Ernährungsforschung
Arthur-Scheunert-Allee 114
14558 Potsdam-Rehbrücke
Professor Dr. Frauke Melchior
Universitätsklinikum der Georg-August-Universität
Göttingen
Zentrum für Biochemie und Molekulare Zellbiologie
Humboldt-Allee 23
37073 Göttingen
Professor Dr. Herbert Waldmann
Max-Planck-Institut für Molekulare Physiologie
Otto-Hahn Str. 11
44202 Dortmund
The Scientific Advisory Board of the FMP formally constituted
itself at its meeting on November 6, 1992. At the
board meeting on November 16, 2000 Professor Reinhard
Jahn was elected chairman.
Der Wissenschaftliche Beirat des FMP konstituierte sich
anläßlich seiner Sitzung am 6. November 1992. Auf der Sitzung
am 16. November 2000 wurde Professor Dr. Reinhard
Jahn zum Vorsitzenden gewählt.

The FMP undertakes research in the field of molecular
pharmacology. In accordance with this objective, the FMP
strives to make contributions of fundamental significance.
In this context, it is engaged in elucidating the structure,
functions and interactions of proteins and in developing
new concepts relating to their pharmacological impact.
Thus, the research activity of the FMP is at the forefront of
drug development. The FMP pursues an interdisciplinary
research approach, which has brought together cellular
signal transduction/molecular genetics, structural biology
and chemical biology. Characteristic for the scientific
work at the FMP is the close interrelationship of chemistry
and biology.
MILESTONES 2003/2004
The years 2003/4 were characterized by the expansion of
the chemistry department. With the appointment of Jörg
Rademann as Professor for Medicinal Chemistry (joint
appointment by the FMP and the Free University Berlin), a
new important area of competence, combinatorial chemi-
FOREWORD
Das FMP betreibt Forschung auf dem Gebiet der Molekularen
Pharmakologie. Dabei ist es bestrebt, Beiträge von
grundsätzlicher Bedeutung zu erbringen. In diesem Rahmen
versucht es, die Struktur, Funktionen und Interaktionen
von Eiweißen (Proteinen) aufzuklären und neue Konzepte
zu ihrer pharmakologischen Beeinflussung zu
entwickeln. Damit ist die Forschungstätigkeit des FMP im
Vorfeld der Entwicklung von Arzneimitteln angesiedelt.
Das FMP verfolgt einen interdisziplinären Forschungsansatz,
der zur Zusammenführung von Zellulärer Signaltransduktion/Molekularer
Genetik, Strukturbiologie und Chemischer
Biologie geführt hat. Kennzeichnend für die
wissenschaftliche Arbeit am FMP ist die enge Verknüpfung
von Chemie und Biologie.
MEILENSTEINE 2003/2004
Die Jahre 2003/2004 waren durch den Ausbau der Chemie
geprägt. Durch die Berufung von Jörg Rademann auf eine
Professur für Medizinische Chemie (gemeinsame Berufung
von FMP und Freier Universität Berlin) wurde neben
der Peptidchemie ein neuer Schwerpunkt „Kombinatorische
Chemie“ geschaffen. Gleichzeitig wurde eine
stry, was established alongside peptide chemistry. At the
same time a Screening Unit was built up, making substance
libraries and screening technology available. Jens
Peter von Kries could be recruited to direct it. The FMP is
thus the first academic institution in Germany which, using
high throughput methods, can identify small molecules
that bind to specific proteins and develop a biological
effect (pages 92-95). These small molecules represent
important tools for research. At the same time they can
also be regarded as prototypes of novel pharmaceuticals.
FMP scientists are centrally involved in building up a new
national network (ChemBioNet; www.chembionet.de),
through which academic groups can obtain access to substance
libraries and screening technology.
With the establishment of the research group Solid State
NMR and the appointment of group leader Bernd Reif as
Professor for Drug Design (joint appointment of the FMP
and the Charité – University Medicine Berlin), the staff of
the NMR Center of the FMP was greatly expanded. An outstanding
event was also the first-time operation of the 900
MHz spectrometer, which represents the current state-of-
Screening Unit aufgebaut, die über Substanzbibliotheken
und Screening-Technologie verfügt. Als Leiter konnte
Jens Peter von Kries gewonnen werden. Damit ist das
FMP die erste akademische Einrichtung in Deutschland,
die kleine Moleküle, die an bestimmte Proteine binden und
eine biologische Wirkung entfalten, im Hochdurchsatzverfahren
identifizieren kann. Diese kleinen Moleküle stellen
wichtige Werkzeuge für die Forschung dar. Zugleich sind
sie aber auch als Prototypen neuartiger Pharmaka zu
betrachten. Wissenschaftler des FMP sind auch in zentraler
Weise am Aufbau eines nationalen Netzwerks (Chem-
BioNet; www.chembionet.de) beteiligt, durch das akademische
Gruppen Zugang zu Substanzbibliotheken und
Screening-Technologie erhalten sollen.
Mit der Einrichtung der Arbeitsgruppe Festkörper-NMR
und der Berufung eines Leiters, Bernd Reif, auf eine Professur
für Drug Design (gemeinsame Berufung von FMP
und Charité – Universitätsmedizin Berlin) wurde das NMR-
Zentrum des FMP personell stark erweitert. Ein herausragendes
Ereignis war auch die Inbetriebnahme eines
900 MHz-Spektrometers, das den aktuellen Stand der Entwicklung
repräsentiert. Weltweit sind erst wenige Geräte
dieses Typs verfügbar. Die erforderlichen Mittel (insge-
3 Foreword

the-art development. Worldwide only a few devices of this
type are in existence. The necessary funding (totaling 4.9
million €) was provided by the BMBF (Federal Ministry of
Education and Research) in the framework of a project
grant and the Berlin Senate Department for Science,
Research and Culture (EFRE funds). Thus an outstandingly
well-equipped and well-staffed center for structural biology
oriented NMR spectroscopy has been created that
has an international reputation.
In June 2003 at a ceremony which included the governing
mayor of Berlin, the cornerstone was laid for the new
Medicinal Genomics Building. It is a joint initiative of the
FMP and the Max Delbrück Center for Molecular Medicine
(MDC). The striking building, designed by the Staab architectural
bureau, provides lab and office space for a staff of
120. The move into the new building will take place in
autumn 2005. Of the 230 employees on the staff of the FMP,
about 40 will work in the new building.
FOREWORD
samt 4,9 Mio €) wurden vom BMBF im Rahmen der Projektförderung
und der Berliner Senatsverwaltung für Wissenschaft,
Forschung und Kultur (EFRE-Mittel) zur Verfügung
gestellt. Damit ist ein gerätemäßig und personell
hervorragend ausgestattetes, international ausstrahlendes
Zentrum für die strukturbiologisch orientierte NMR-
Spektroskopie etabliert.
Im Juni 2003 wurde gemeinsam mit dem Max-Delbrück-
Centrum für Molekulare Medizin (MDC) im Beisein des
Regierenden Bürgermeisters von Berlin der Grundstein für
den Neubau „Medizinische Genomik“ gelegt. Das markante,
von dem Architektenbüro Staab entworfene Gebäude
bietet 120 Mitarbeitern Labor- und Büroarbeitsplätze.
Der Bezug des neuen Gebäudes wird im Herbst 2005 erfolgen.
Von den 230 Mitarbeitern des FMP werden ca. 40 im
neuen Gebäude tätig sein.
PERSPEKTIVEN
Arzneimittel entfalten ihre Wirkung durch Interaktion ihrer
Inhaltsstoffe (Wirkstoffe) mit körpereigenen Strukturen.
Bei diesen Andockstellen im Organismus (Zielstrukturen,
„targets“) handelt es sich fast ausnahmslos um Proteine
(> 99%). Bisher ist erst ein sehr kleiner Teil der Eiweiße des
menschlichen Körpers als Zielstruktur für einen Wirkstoff
erschlossen (ca. 500). Sicher kommen nicht alle 100.000
Eiweiße des Organismus als „targets“ für Wirkstoffe in
Frage. Man geht aber davon aus, dass zumindest einige
tausend geeignet sind. Damit stellt sich die Aufgabe, die
derzeit schmale Basis der Arzneimitteltherapie durch
Identifizierung neuer Zielstrukturen zu erweitern und so
bestehende Therapiekonzepte zu verbessern und neue zu
etablieren. Hier sieht das FMP ein zentrales Tätigkeitsfeld.
Zu den Proteingruppen, die gegenwärtig am FMP bearbeitet
werden, gehören als klassische „targets“ Membranproteine
wie Rezeptoren und Kanalproteine, aber auch
Proteine, die bisher kaum als „targets“ in Betracht gezogen
wurden. Zu dieser Gruppe gehören Transkriptionsfaktoren,
die Gene regulieren, Interaktionsdomänen von
Proteinen und sogenannte Ankerproteine, die Proteine in

PERSPECTIVES
Drugs achieve their effect through interaction of their contents
(substances) with the body’s own structures. At
these docking sites in the organism (targets), this almost
without exception involves proteins (> 99%). Until now,
drugs have been developed for only a small number of the
proteins of the human body (approx. 500).
To be sure, not all 100,000 proteins of the organism are
appropriate as a target for substances. However, it is
assumed that at least several thousand are suitable. Therefore,
the task is to expand the present narrow base of
drug therapy by identifying new target structures and thus
to improve existing therapy concepts and establish new
ones. Here the FMP sees a central field of activity. The protein
groups that are currently being processed at the FMP
include as classical targets membrane proteins such as
receptors and channel proteins, but also proteins that up
till now have hardly been considered targets. To this group
belong transcription factors, which regulate genes, interaction
domains of proteins and so-called anchor proteins,
der Zelle zu größeren funktionellen Komplexen zusammenfügen,
sowie extrazelluläre Fibrillen. (die z. B. bei der Alzheimerschen
Erkrankung vermehrt auftreten).
Pharmazeutische Firmen ziehen sich immer mehr aus der
Forschung im Vorfeld eines „proof of principle“, d. h. aus
der „high-risk“ Forschung, die der Arzneimittelentwicklung
vorausgeht, zurück. Außerdem ist seit langem eine
Verlagerung der Forschungsabteilungen pharmazeutischer
Firmen ins Ausland zu beobachten. Durch die Stärkung
der chemischen Forschung und besonders durch die
Einrichtung der Screening Unit hat das FMP seine Kompetenz
innerhalb der Wertschöpfungskette, die zu neuen
Arzneimitteln führt, erheblich in Richtung Anwendung
erweitert, ohne allerdings seine feste Verankerung in der
Grundlagenforschung aufzugeben. Es ist kann nun Vorschläge
zu neuen „targets“ mit Vorschlägen zu Wirkstoffkandidaten
verbinden und evtl. auch eine erste pharmakologische
Charakterisierung potentieller „targets“
präsentieren. Wir hoffen, dass die Erweiterung der Plattform
dazu beiträgt, die Lücke zwischen pharmakologischer
Grundlagenforschung in den Forschungseinrichtungen
und der Arzneimittelentwicklung in der
pharmazeutischen Industrie zu schließen und damit das
which assemble the proteins in the cell to larger functional
complexes, as well as extracellular fibrils (which e.g.
are increasingly observed with Alzheimer’s disease).
Pharmaceutical companies are withdrawing more and
more out of research preparatory to a “proof of principle“,
i.e. from “high-risk” research which precedes drug development.
Moreover, for a long time it has been observed
that the research departments of pharmaceutical companies
have been relocating abroad. Due to the strengthening
of chemical research and particularly due to the establishment
of the Screening Unit, the FMP has considerably
expanded its competence within the value creation chain
that leads to new drugs with regard to application, yet
without relinquishing its firm anchoring in basic research.
Now it can combine suggestions for new “targets” with
suggestions for substance candidates and possibly present
a first pharmacological characterization of potential
“targets”. We hope that the extension of the platform will
help close the gap between pharmacological basic
research in the research institutes and drug development
in the pharmaceutical industry, and thus that the environ-
Umfeld für Unternehmen des Pharma-Branche attraktiver
zu gestalten. Diesem Ziel sollen auch die verschiedenen
Netzwerkinitiativen dienen, an denen das FMP maßgeblich
beteiligt ist. Beispiele sind die Etablierung eines Netzwerks
„Arzneimittelentwicklung in Berlin und Brandenburg“
und die bereits erwähnte nationale Initiative
„ChemBioNet“.
Wie in den vergangenen Jahren stehen Beiträge der
Arbeitsgruppen über ihre wissenschaftliche Arbeit im Zentrum
dieses Berichtes 2003/2004. Er enthält aber auch die
Leistungsbilanz für den Berichtszeitraum. So wird die
Publikationstätigkeit, die Einwerbung von Drittmitteln und
die Beteiligung des FMP an der Ausbildung von Studierenden
verschiedener Disziplinen dargestellt.
Der Erfolg der letzten Jahre ist den Mitarbeiterinnen und
Mitarbeiter des FMP zuzuschreiben, bei denen ich mich
herzlich für ihr Engagement bedanken möchte. Mein Dank
richtet sich auch an den wissenschaftlichen Beirat, der
das FMP konstruktiv-kritisch begleitet hat.
5 Foreword

ment for companies in the pharma sector will become
more attractive. Various network initiatives in which the
FMP plays a major role shall also serve this goal. Examples
are the establishment of a network “Drug Development
in Berlin und Brandenburg” and the already mentioned
national initiative “ChemBioNet”.
As in past years, the contributions of the various research
groups about their scientific work stand in the center of
this research report 2003/2004. However, it also contains
a performance account for the period of the report. Thus,
publication activity, solicitation of third-party funds and the
participation of the FMP in the education and training of
students of the various disciplines are also presented.
The success of the last years is due to the colleagues of
the FMP, whom I would like to thank very much for their
dedication. My thanks also go to the Scientific Advisory
Board for its constructive and critical support of the FMP.
FOREWORD
I would like to invite readers to take an informative and
interesting tour through the pages of this report to see how
the FMP has developed since its founding 12 years ago.
Walter Rosenthal, Director
Ich wünsche den Lesern einen aufschlussreichen und
interessanten Streifzug durch das FMP, wie es sich
12 Jahre nach seiner Neugründung darstellt.
Walter Rosenthal, Direktor
April 2005

CONTENTS / INHALT
Scientific Advisory Board / Wissenschaftlicher Beirat ......................................................................................................... 2
Foreword / Vorwort ..................................................................................................................................................................... 3
Section Structural Biology / Bereich Strukturbiologie
Introduction ......................................................................................................................................................................... 10
Protein Structure ................................................................................................................................................................ 12
Solution NMR....................................................................................................................................................................... 16
Structural Bioinformatics .................................................................................................................................................. 20
Molecular Modelling .......................................................................................................................................................... 24
Solid State NMR ................................................................................................................................................................. 28
Protein Engineering ............................................................................................................................................................ 31
Section Cellular Signalling / Molecular Genetics /
Bereich Signaltransduktion/ Molekulare Genetik
Introduction ......................................................................................................................................................................... 36
Protein Trafficking .............................................................................................................................................................. 39
Anchored Signalling ........................................................................................................................................................... 42
Cellular Imaging .................................................................................................................................................................. 46
Molecular Cell Physiology ................................................................................................................................................ 49
Biochemical Neurobiology ................................................................................................................................................ 52
Biophysics ........................................................................................................................................................................... 56
Cytokine Signaling .............................................................................................................................................................. 59
Molecular Myelopoiesis .................................................................................................................................................... 62
Mouse Models..................................................................................................................................................................... 64
Cellular Signal Processing ................................................................................................................................................ 65
Section Chemical Biology / Bereich Chemische Biologie
Introduction ......................................................................................................................................................................... 70
Peptide Synthesis ............................................................................................................................................................... 72
Peptide Lipid Interaction/Peptide Transport .................................................................................................................. 75
Peptide Biochemistry ......................................................................................................................................................... 80
Mass Spectrometry ............................................................................................................................................................ 83
Synthetic Organic Biochemistry ...................................................................................................................................... 86
Medicinal Chemistry .......................................................................................................................................................... 89
Screening Unit .................................................................................................................................................................... 92
Scientific and technical services / Wissenschaftlicher und technischer Service
Microdialysis ....................................................................................................................................................................... 98
DNA Sequencing .............................................................................................................................................................. 101
Public Relations and the Media / Presse- und Öffentlichkeitsarbeit ........................................................................ 102
Administration / Verwaltung ........................................................................................................................................... 106
Computer Services / Computer-Service ........................................................................................................................ 108
Contents
7

Flip-book
Daumenkino
Appendix / Anhang
Peer reviewed Articles 2003 / Originalarbeiten ............................................................................................................ 110
Peer reviewed Articles 2004 / Originalarbeiten ............................................................................................................ 114
Reviews 2003/2004 / Übersichtsarbeiten ...................................................................................................................... 121
Contributions in Monographs 2003/2004 / Beiträge zu Sammelwerken ....................................................................121
Monographs 2003/2004 / Monographien ...................................................................................................................... 122
Memberships in Editorial Boards 2003/2004 / Mitgliedschaften in Editorial Boards ..............................................123
Invited Talks 2003/2004 / Eingeladene Vorträge .......................................................................................................... 123
External funding 2003/2004 / Drittmittel.......................................................................................................................... 128
Participation in Research Networks 2003/2004 / Beteiligung an Netzwerken und Verbundprojekten ................ 130
Cooperations with contract 2003/2004 / Vertragliche Kooperationen....................................................................... 132
Meetings, Workshops, Symposia 2003/2004 / Wissenschaftliche Veranstaltungen ............................................... 134
Work in Panels 2003/2004 / Gremienarbeit.................................................................................................................... 135
Review Activities 2003/2004 / Gutachtertätigkeit ......................................................................................................... 135
Academic Teaching 2003 / 2004 / Lehre.......................................................................................................................... 138
Calls for Appointments 2003/2004 / Rufe........................................................................................................................ 141
Post-Doctoral Lecture Qualifications 2003/2004 / Habilitationen .............................................................................. 141
Graduations 2003/2004 / Promotionen ........................................................................................................................... 142
Diploma theses 2003/2004 / Diplomarbeiten ................................................................................................................. 143
Internships 2003/2004 / Praktikanten ............................................................................................................................. 144
Guest Scientists 2003/2004 / Gastwissenschaftler ...................................................................................................... 148
Lectures at the FMP 2003 / Kolloquien und Seminare am FMP .................................................................................. 149
Lectures at the FMP 2004 / Kolloquien und Seminare am FMP ................................................................................. 153
Technology Transfer 2003/2004 / Technologietransfer ................................................................................................ 156
Structure of the Forschungsinstitut für Molekulare Pharmakologie (FMP) / Organigramm .................................. 158
Index / Namensregister ................................................................................................................................................... 160
Maps / Lagepläne ............................................................................................................................................................. 164

STRUCTURAL BIOLOGY

SECTION STRUCTURAL BIOLOGY
Prof. Hartmut Oschkinat
Department Head: NMR-supported Structural Biology
(Secretary: Andrea Steuer)
The research themes of the section cover structural and
functional studies of receptors and proteins involved in
intracellular signalling, and projects on the biophysical
description of protein aggregate formation. Strong emphasis
is thereby put on a structural characterization of membrane-integrated
receptors, channels and fibril-forming
systems, in combination with exploring the potential of
solid-state magic angle spinning NMR for studying such
systems. As a special theme with regard to intracellular
signalling, non-catalytical protein domains that mediate
interactions with linear peptide segments in target proteins
are analyzed with respect to specificity and affinitydetermining
sequence features. A growing part of the
research is the systematic derivation of small organic
INTRODUCTION
BEREICH STRUKTURBIOLOGIE
Prof. Hartmut Oschkinat
Abteilungsleiter: NMR-unterstützte Strukturforschung
(Sekretariat: Andrea Steuer)
Der Bereich Strukturbiologie erforscht Struktur und Funktion
von Rezeptoren und Proteinen, die an der intrazellulären
Signalübertragung beteiligt sind, sowie die Entstehung
von Proteinaggregaten. Im Mittelpunkt des
Interesses steht die strukturelle Charakterisierung von
Rezeptoren, Kanälen und fibrillenbildenen Systemen in
Kombination mit der Erforschung des Potentials der
„magic-angle-spinning“ Festphasen-NMR für das Studium
solcher Systeme. Im Hinblick auf die intrazelluläre Signalübertragung
wird besonderes Augenmerk auf die spezifitäts-
und affinitätsbestimmenden Eigenschaften nichtkatalytischer
Proteindomänen gelegt. Diese vermitteln die
Interaktionen mit linearen Peptidsegmenten in Zielproteinen.
Zunehmend rückt die systematische Derivatisierung
kleiner organischer Moleküle als Modulatoren von Protein-Protein-Interaktionen
und der Amyloid-Bildung in den
Mittelpunkt des Interesses. Wichtige inhaltliche Brücken
compounds as modulators of protein-protein interactions
and amyloid formation. There are strong links to the
screening facility of the FMP through the molecular
modellling and NMR aspects of small-molecule inhibitor
development.
The department hosts expertise in the areas of structural
bioinformatics, molecular biology, solution and solid-state
NMR spectroscopy, and molecular modelling. NMR
spectroscopy is used as the major tool for determining
structures of biological macromolecules in solution, and
of quasi-solid preparations which are difficult to crystallize,
like amyloid fibrils and membrane proteins in nativelike
lipids. It is particularly well suited for studying the
dynamics of protein structures and interactions with very
weakly binding ligands. The various skills are contained in
currently six research groups, Protein Engineering
(Christian Freund), Molecular Modelling (Ronald Kühne),
Structural Bioinformatics (Gerd Krause), Protein Structure
(Hartmut Oschkinat), Solid State NMR (Bernd Reif) and
zum Screening-Labor des FMP sind die Molekülmodellierung
und NMR-Aspekte der Entwicklung niedermolekularer
Hemmstoffe.
Der Bereich kann Expertise auf den Gebieten strukturelle
Bioinformatik, Molekularbiologie, Flüssigkeits- und Festkörper-NMR-Spektroskopie
sowie Molekülmodellierung
vorweisen. Hauptsächlich mittels NMR-Spektroskopie
werden Strukturen biologischer Makromoleküle in Lösung
und in Präparationen an der Festkörperphase bestimmt
(Amyloidfibrillen und Membranproteine in natürlichen
Lipiden). NMR-Spektroskopie ist besonders geeignet für
Studien zur Dynamik von Proteinen und Interaktionen mit
sehr schwach bindenden Liganden. Gegenwärtig sechs
Arbeitsgruppen bieten wissenschaftliche Kompetenz in
Protein Engineering (Christian Freund), Molekülmodellierung
(Ronald Kühne), Struktureller Bioinformatik (Gerd
Krause), Proteinstruktur (Hartmut Oschkinat), Festkörper-
NMR (Bernd Reif) und Lösungs-NMR (Peter Schmieder).
Der Bereich ist mit einer Vielzahl von Lösungs- und Festkörper-NMR-Spektrometern
(400 bis 900 MHz) sowie Geräten
zur analytischen Ultrazentrifugation und isothermischen
Kalorimetrie ausgestattet. In den zurückliegenden

Solution NMR (Peter Schmieder). The instrumentation
includes a variety of solution and solid-state NMR spectrometers
ranging from 400 to 900 MHz, and equipment for
analytical ultracentrifugation and isothermal calorimetry.
In the past two years, the direction of research has been
shifted stronger towards solid-state NMR and its applications,
resulting in the installation of a new group focused
on solid-state NMR and amyloid-forming systems. Furthermore,
a 700 MHz wide bore NMR spectrometer was purchased.
zwei Jahren hat sich der Fokus der Forschung stärker in
Richtung Festkörper-NMR und deren Anwendungen verschoben.
Dies führte zur Ansiedlung einer neuen Arbeitsgruppe,
die sich mit Festkörper-NMR und Amyloid-bildenden
Systemen befasst. Außerdem wurde ein 700
MHz-„wide bore“-NMR-Spektrometer in Betrieb genommen.
11 Structural Biology

PROTEIN STRUCTURE
Group Leader: Prof. Hartmut Oschkinat
STRUCTURAL CHARACTERIZATION OF
PROTEIN-PROTEIN-INTERACTIONS
The main focus of our group is the structural characterization
of protein-protein interactions responsible for the
reception and transduction of signals in biological
systems. This research concerns protein domains which
recognise specific peptides and on the long run investigations
on membrane integrated receptors and receptorligand
complexes. It is embedded into efforts towards
structural genomics of soluble and membrane proteins
and supported by NMR-technical developments. The
technical developments aim at higher throughput and
towards concepts for structure determination of membrane
proteins by solid-state NMR.
Structure and function of non-catalytic protein
domains
Non-catalytic protein domains mediate protein-protein
interactions through the recognition of peptide segments
in a highly specific manner. Such interactions govern the
reversible assembly of macromolecular signalling complexes
which finally form a logical network of interacting proteins,
thereby transmitting information. Our investigations
on non-catalytic protein domains typically involve a combination
of structural and functional studies, e.g. by using
peptide libraries, NMR spectroscopy and a variety of other
biophysical techniques, aiming at an understanding of
molecular recognition and drug design.
We have determined the structures of a number of WW
domain-peptide complexes, and the structure of an SH3
domain in complex with a canonical and a non-canonical
peptide revealing two different binding sites. Intense studies
with PDZ and WW domains involving NMR and peptide
library screens where set out to derive structure-activity-relationships
and to provide data for a theoretical
analysis of domain specificity. We quantitatively determined
the specificity profiles of three representative PDZ
domains from the AF6, ERBIN and SNA1 proteins, with
respect to the complete ligand sequence space of C-terminal
peptides. This quantification was achieved by combining
efficiently dissociation constant measurements and
statistical analysis of synthetic peptide libraries, using an
Analysis of Variance (ANOVA) approach. Predicted in vitro
affinities were validated by designing super-binding peptides
which indeed revealed the lowest Kd values and the
highest PDZ domain specificity. The coverage of the com-
plete ligand sequence space for the three PDZ domains
made it feasible to compare their ligand selectivity and the
overlap of their recognized ligand sequence subspaces.
NMR structure determination in a structural
genomics context
As a part of the protein structure factory project, a number
of protein structures were generated, including those
of the subunit B8 of complex I, of the p47 SEP domain, and
of a BRCT domain.
The solution structure of the subunit B8 from ubiquinone
oxidoreductase (Complex I) (CI-B8) shows a thioredoxin
fold (Fig. 1) with remarkable similarities to thioredoxin-like
Fe2S2-ferredoxins. A detailed comparison to these proteins
show remarkably high similarities of surface properties
and possibly catalytically active residues. The redoxpotential
of the disulfide-bond is comparable to that of the
catalytically active disulfides in thioredoxin-like proteins.
This led to the hypothesis that this subunit may be involved
in the regulation of complex I.
The BRCT domain occurs in a number of different signal
transduction proteins involved in transcriptional regulation,
DNA repair, cell cycle progression and cancer suppression.
The C-terminal region of the breast cancer associated
tumor suppressor protein BRCA1 consists of two
successive BRCT domains, separated by a short linker. We
have, in collaboration with the group of Udo Heinemann at
the MDC, Berlin-Buch, solved the solution structure of
BRCT-c, the C-terminal BRCT domain of BRCA1. Analysis
of the structure and comparison with other members of the
BRCT superfamily has revealed that only a small number of
conserved residues are necessary for the formation and
stabilization of the BRCT motif.
The structure of the SEP domain of the protein p47 was
determined and a hypothesis as to its function derived and
verified. A particular feature of this fold is the conservation
of residues in two loops, and of aromatic residues very
close to it (Fig. 2). These loops are remarkably rigid, and
the upper part of the structure in Fig. 2 resembles cysteine
protease inhibitors like cystatins and stefins. An assay
showed a weak inhibition of cathepsin L by the p47 SEP
domain.
Toward structure determination of membrane
proteins by NMR
Major forces were concentrated on the further development
of a structure determination concept for proteins
using magic-angle spinning solid-state NMR. Such a
method would be very welcome for systems which are

FIGURE 1
Ribbon diagram of subunit B8 (left) from ubiquinone oxidoreductase (Complex I, right)
difficult to crystallize and out of range for solution NMR,
like membrane proteins, amyloids, and structure forming
proteins like actin filaments, for example. Along with
various membrane proteins, a microcrystalline sample of
the α-spectrin SH3 domain is studied as a model system
which yields suitable NMR spectra for structure determination.
At the beginning of the study resonance
assignments for all 13C and 15N resonances were achieved,
followed by an assignment of non-exchangeable, and
later also of most exchangeable protons. Based on the
achieved assignments a first structure of a protein was
determined. A key aspect of this study was the generation
of biosynthetically site directed labelled samples by using
either 1,3- 13C-glycerol or 2- 13C-glycerol as carbon sources.
The respective proton driven spin diffusion spectra received
from those samples showed a dramatically increased
number of long-range restrains that were used for structure
calculations. We have now refined the original structure
of the α-spectrin SH3 domain using 889 interresidue
carbon-carbon and 6 15N- 15N restraints, all obtained by
solid-state MAS NMR. The derived procedure should be
widely applicable to small membrane proteins and amyloid
systems if the proteins can be expressed in bacteria.
First steps towards membrane proteins were taken using
the snake toxin neurotoxin II bound to the nicotinic acetyl
choline receptor as a test system. The spectra show
remarkably sharp lines, and an evaluation of the spectra
is under way.
Group members
Dr. Linda Ball
Dr. Anne Diehl
Dr. Katja Fälber
Dr. Jeremy Flinders
Dr. Ludwig Krabben
Dr. Dirk Labudde
Dr. Dietmar Leitner
Dr. Ricardo Pires
Dr. Barth van Rossum
Prisca Boisguerin (Doctoral student)
Christoph Brockmann (Doctoral student)
Federica Castellani (Doctoral student)
Michele Fossi (Doctoral student)
Matthias Hiller (Doctoral student)
Henrik Holtmann (Doctoral student)
Mangesh Joshi (Doctoral student)
Christian Köhler (Doctoral student)
Structural Biology
13

B
Thien-Thi Mac (Doctoral student)
Vivien Lange (Doctoral student)
Doreen Pahlke (Doctoral student)
Ilja Poliakov (Doctoral student)
Nikolaj Schröder (Doctoral student)
Michael Soukenik (Doctoral student)
Carolyn Vargas (Doctoral student)
Urs Wiedemann (Doctoral student)
Stefan Jehle (Student)
Heide Evers (Technical assistance)
Lilo Handel (Technical assistance)
Martina Leidert (Technical assistance)
Kristina Rehbein (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Struktur und Mechanismus der 3,4-Dihydroxy-2-butanon-
4-phosphat-Synthase“ (OS 106/ 4-2)
Hartmut Oschkinat
Deutsche Forschungsgemeinschaft
„Analyse von essentiellen WW-Domänen in ß-sheet-
Strukturen am Beispiel der WW-Domäne mittes Einbaus
nicht natürlicher Aminosäuren“ (OS 106/5-1,5-2)
Hartmut Oschkinat
N
C
FIGURE 2
Structural ensemble of the p47 SEP domain. The residues with side chains displayed in black and green are highly conserved.
N
Deutsche Forschungsgemeinschaft
„Schlüsselreaktionen der biologischen Wasserstoffaktivierung
am Beispiel der [NiFe]-Hydrogenasen“ (Teilprojekt
C1 im Sonderforschungsbereich 498 “Protein-Kofaktor-
Wechselwirkungen in biologischen Prozessen“)
Hartmut Oschkinat, Bärbel Friedrich (Humboldt-Universität
zu Berlin)
Deutsche Forschungsgemeinschaft
„Bestimmung der Raumstrukturen von Rezeptor-gebundenen
Agonisten und Antagonisten mittels Festkörper-NMR-
Spektroskopie“ (Teilprojekt B1 im Sonderforschungsbereich
449 „Struktur und Funktion membranständiger
Rezeptoren“)
Hartmut Oschkinat
Deutsche Forschungsgemeinschaft
„Theoriegestützte NMR-spektroskopische Analyse von
Protein-Ligand-Wechselwirkungen unter Verwendung von
Peptidbibliotheken“ (Teilprojekt in der Forschergruppe 299
„Optimierte molekulare Bibliotheken zum Studium biologischer
Erkennungsprozesse“)
Hartmut Oschkinat, Michael Bienert
C

Deutsche Forschungsgemeinschaft
„Analyse von essentiellen Wechselwirkungen in ß-sheet-
Strukturen am Beispiel der WW-Domäne mittels Einbau
nicht natürlicher Aminosäuren“ (Teilprojekt in der Forschergruppe
475 “Bildung und Stabilität von β-Faltblättern)
Hartmut Oschkinat
Bundesministerium für Bildung und Forschung
„Strukturanalyse mit hohem Durchsatz für medizinisch
relevante Proteine“ (01GG 9812, BMBF-Leitprojekt Proteinstrukturfabrik)
Hartmut Oschkinat
Bundesministerium für Bildung und Forschung
„Aufbau einer Technologieplattform NMR-Messtechnik
für die Proteomforschung“ (0312832)
Hartmut Oschkinat
Bundesministerium für Bildung und Forschung
“Festkörper-NMR-Spektroskopie“ (0312890G, Teilprojekt
im Verbund „Proteomweite Analyse membrangebundener
Proteine – ProAMP“)
Hartmut Oschkinat
Bundesministerium für Bildung und Forschung
„Screening von Substanzbibliotheken und Struktur-basierten
Wirkstoffdesign“ (0312992J, Teilprojekt im Verbundvorhaben
„Strukturproteomik“ – Konsortium Hamburg:
„Hochdurchsatz-Strukturanalyse von Mycobacteriumtuberculosis-Zielproteinen
und ihrer Ligandenkomplexe
zur Suche nach Wirkstoffen“
Hartmut Oschkinat, Jens Peter von Kries
European Community
“Exploiting synthetics SH2-scaffolded repertoire libraries
to profile cancer cells and to interfere with cancer-related
phenotypes” (QLK3-2000-00924)
Hartmut Oschkinat
European Community
„Interactions between growth factors and glycosaminoglycans
in arterial disease” (BMH4-98-3289)
Hartmut Oschkinat
Selected publications (FMP authors in bold)
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Prot Sci 12, 491-500
Kahmann JD, Wecking DA, Putter V, Lowenhaupt K, Kim
YG, Schmieder P, Oschkinat H, Rich A, Schade M (2004)
The solution structure of the N-terminal domain of E3L
shows a tyrosine conformation that may explain its redu-
ced affinity to Z-DNA in vitro. Proc Natl Acad Sci USA 101,
2712-2717
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kühne R, Oschkinat H (2004) The oxidized subunit
B8 from human complex I adopts a thioredoxin fold.
Structure 12, 1645-1654
Gaiser OJ, Ball LJ, Schmieder P, Leitner D, Strauss H,
Wahl M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
Structure, Backbone Dynamics, and Association
Behavior of the C-Terminal BRCT Domain from the Breast
Cancer-Associated Protein BRCA1. Biochemistry 43,
15983-15995
Collaborations
Adelbert Bacher, Garching
Alan Fersht, FRS, Cambridge, UK
Paul Gooley, Melbourne
Robert G. Griffin, Cambridge, USA
Udo Heinemann, Berlin
Klaus-Peter Hofmann, Berlin
Dieter Oesterhelt, Martinsried
Michael Nilges, Paris
Jens Schneider Mergener, Berlin
Rudolf Volkmer-Engert, Berlin
Structural Biology
15

SOLUTION NMR
Group Leader: Dr. Peter Schmieder
APPLICATIONS OF SOLUTION STATE NMR
TO ADDRESS BIOLOGICALLY IMPORTANT
QUESTIONS
Peter Schmieder, Holger Strauss, Christian Appelt, Janina
Hahn, Brigitte Schlegel
NMR spectroscopy is an important technique to address
numerous questions regarding the structure of various
kinds of molecules, including their three-dimensional
structure. Solution-state NMR is well established as a tool
to obtain the three-dimensional structure at atomic resolution
for soluble biomolecules. Solid-state NMR is
currently emerging as a technique to obtain structural
information, where solution-state NMR is becoming
exceedingly difficult, namely in the case of membrane proteins.
NMR spectroscopy, preferably in solution, can also provide
valuable information on the interaction of proteins and
ligands, either in the case of a particular ligand of interest
or in the case of large libraries used in screening protocols.
Last but not least, NMR is extremely important for the
determination of the constitution of smaller molecules.
In our group we use the full repertoire of solution-state
NMR techniques in conjunction with a variety of labeling
patterns to address questions of biological and pharmacological
importance. These range from the development
of new techniques for solution-state NMR to the elucidation
of the constitution of biologically active peptides and
the determination of the three-dimensional structure of
peptides and proteins. In addition, NMR spectroscopy is
offered as a tool to researchers and companies throughout
the Berlin area.
Structure of the chromophore-binding pocket of
the cyanobacterial phytochrome Cph1
Phytochrome photoreceptors control numerous developmental
processes in all plants. Light is absorbed by a covalently
bound tetrapyrrole chromophore, phytochromobilin
(PΦB), which is incorporated autocatalytically into a chromophore-binding
domain in the N-terminal sensory module
of the protein. The physiological function of phytochrome
is associated with the photochemical production of the
thermodynamically stable P fr signaling state following light
absorption by the P r- ground state (λ max ~660 nm). The P fr
form itself is photoactive (λ max ~730 nm), quantum absorption
leading to the production of P r . In daylight the photocycle
establishes a dynamic equilibrium with a constant
level of P fr . The difference between the P r and P fr is attributed
to the Z-E-isomerization of a double bond within the
chromophore, as suggested from NMR data from isolated
chromopeptides.
With the discovery of Cph1 in the cyanobacterium Synechocystis
sp. PCC 6803 it was realized that phytochromes
are also present in prokaryotes. The native chromophore
of Cph1 in Synechocystis as well as in many other cyanobacterial
phytochromes is phycocyanobilin. Subsequently,
phytochromes have also been detected in other cyanobacteria
and even non-photosynthetic bacteria, where the
chromophore frequently is biliverdine. Heterologous expression
of phytochromes in E. coli yields apoproteins that
are usually capable of self-assembling with the chromophore
to become red/farred photochromic holoproteins. A
deeper understanding of the photochromic mechanism of
phytochrome proteins requires detailed knowledge of the
structure of the chromophore pocket. No structural information
at atomic resolution, however, is available to date.
We want to use solution state NMR spectroscopy to obtain
structural information on the chromophore binding pocke
in the two parent states (P r and P fr). To separate the signals
of the chromophore from those of the protein, deuteration
of the protein is one approach. Since the chromophore is
relatively small compared to the protein, labeling of PCB
with nitrogen and carbon is also necessary. While the production
of proteins in isotopically-labeled form is a wellestablished
procedure and while the same is also possible
with DNA and RNA, the production of other types of
molecules enriched with 13 C and/or 15 N is usually prohibitively
expensive. In the case of the PCB chromophore
necessary for the present investigation, the problem is
somewhat ameliorated by the fact that PCB is the chromophore
in the light-harvesting antenna of photosynthetic
bacteria and that the phycobilisome which contains the
chromophore molecules represents a major fraction of the
soluble proteins in cyanobacteria. We therefore decided
to produce isotopically-labeled PCB using Synechocystis
sp. PCC 6803 itself (Figure 1).
Using the different labeling pattern, it was possible to
obtain NMR spectra of sufficient quality to extract structural
information. By recording one-dimensional 15 N-spectra,
we could solve the much-discussed question whether the
chromophore is protonated in the two parent states, which
has strong implications on the reactions taking place in the
phytochrome photocycle. The nitrogen chemical shifts
obtained indicate that both the P r and the P fr form are in
fact protonated. Using carbon-labeled PCB, three-dimensional
edited NOESY spectra could be obtained that will
yield an assignment of the chromophore resonances and
will allow the selection of a most likely structure from

FIGURE 1
Preparation of 15N and 13C, 15N-labeled Phycocyanobilin (PCB) from Synechocystis sp. PCC 6803: The cyanobacteria are grown on the appropriately
labeled medium (a). After harvesting the cells, the phycobilisome is separated by a sucrose density centrifugation (b). Finally, PCB is extracted
using methanol (c).
several structural models. An analysis of those spectra is
still in progress, a next step towards a structure of the
chromophore-binding pocket is the introduction of protonated
amino acids into a deuterated background to detect
interactions between the chromophore and amino acids
residues lining the binding pocket.
Structure of antimicrobial peptides in a membrane-mimicking
environment
Antimicrobial peptides are part of the natural immune
system of many living organisms. A broad range of microorganisms
that involves gram-positive, gram-negative
bacteria and fungi is affected by these peptides, which are
evolutionary ancient weapons. Usually a cocktail of multiple
peptides is present, supplementing the pathogen-specific
immune response, which occurs relatively slowly.
Since bacterial resistance to existing antibiotics is constantly
increasing, these peptides have gained in interest
since they have the potential to form an entirely new drug
generation.
Despite a growing interest in this type of peptides, their
mechanism of action is largely unknown. Since a replacement
of L-amino acids with their D-enantiomers does not
usually alter the antimicrobial activity of the peptides
unless the overall structure is disrupted, a mechanism via
a specific protein receptor is unlikely. It is rather the bacterial
membrane that is the most likely target of the antimicrobial
assault. The outer membrane leaflets of grampositive
as well as gram-negative bacteria are negatively
charged, consisting mainly of phosphatidylglycerol or lipopolysaccharide,
respectively. In contrast, the outer leaflet
of mammalian cell membranes contains mostly phosphatidylcholine
and is thus charge neutral at physiological pH.
It is assumed that the negative charge is responsible for
the selectivity.
In order to gain insight into the mechanism of action of
antimicrobial peptides at the atomic level, we have determined
the structure of the cyclic, cationic antimicrobial
peptide cyc-(RRWWRF) free in aqueous solution and
bound to detergent micelles using NMR spectroscopy. The
peptide shows a rather flexible but nevertheless ordered
structure in water, but a distinct structure is formed when
the peptide is bound to a detergent micelle. The structures
in the neutral and negatively charged micelles are
nearly identical, resembling two β-turns. The orientation
of the amino acid side chains creates an amphipathic
molecule with the peptide backbone forming the hydrophilic
part. The orientation of the peptide in the micelle was
Structural Biology
17

determined using NOEs and the accessibility of the peptide
from outside the micelle as probed using paramagnetic
agents. The peptide is oriented mainly parallel to the
micelle surface in both detergents. Substitution of the arginine
and tryptophan residues is known to influence the
antimicrobial activity. Therefore, the structure of the micelle-bound
analogs cyclo(RRYYRF), cyclo(KKWWKF) and
cyclo(RRNalNalRF) were determined, having remarkable
similarities in backbone conformation and side-chain orientation.
Based on these structures, molecular dynamics
simulations of the peptide in an explicit membrane at
various peptide-to-lipid ratio were performed. We observe
that the NMR structure of the peptide is stable also after
100 ns simulation. At a peptide-to-lipid ratio of 2/128, the
membrane is only little affected compared to a pure DPPC
lipid membrane, but at a ratio of 12/128, the water-lipid
interface becomes more fuzzy, the water molecules can
reach deeper into the hydrophobic core, and the water
penetration free energy barrier changes. Moreover, we
observe that the area per lipid decreases and the deuterium
order parameters increase in the presence of the
peptide. We suggest that the changes in the hydrophobic
core together with the changes in the head groups result
in an imbalance of the membrane-stabilizing forces that
will lead to the disruption of the membrane.
FIGURE 2
Structure of the cyclic peptide cyc-(RRWWRF) bound to DPC micelles. The molecule
exhibits an amphipatic structure with the peptide backbone as the hydrophilic
part (blue) and the aromatic side chains as the lipophilic part (brown). The
orientation of the peptide has been determined to be parallel to the micelle surface
with the longest axis, with the lipophilic side chains penetrating into the
micelle.
Group members
Holger Strauss (Doctoral student)
Christian Appelt (Doctoral student)
Janina Hahn (Doctoral student)*
Brigitte Schlegel (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„NMR-spectroscopic investigation of light-induced structural
changes in protein-chromophore complexes” (Teilprojekt
B6 im Sonderforschungsbereich 498 „Protein-
Kofaktor-Wechselwirkungen in biologischen Prozessen“
Peter Schmieder
Fonds der Chemischen Industrie
„NMR-spektroskopische Strukturuntersuchung von photoschaltbaren
Peptidliganden für die Numb-PTB-Domäne
sowie von PTB/Ligandkomplexen“
Kekulé-Stipendium
Christian Appelt (Promotionsstipendium)
* part of period reported

Selected publications (FMP authors in bold)
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Prot Sci 12, 491-500
Strauss H (2003) A device for facilitating the use of the
French Press. Anal Biochem 321, 276-277
Hupfer M, Rübe B, Schmieder P (2004) Origin and diagenesis
of polyphosphate in lake sediments: A 31 P-NMR
study. Limnol Oceanogr 49, 1-10
Kahmann JD, Wecking DA, Putter V, Lowenhaupt K, Kim
YG, Schmieder P, Oschkinat H, Rich A, Schade M (2004)
The solution structure of the N-terminal domain of E3L
shows a tyrosine conformation that may explain its reduced
affinity to Z-DNA in vitro. Proc Natl Acad Sci USA 101,
2712-2717
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kühne R, Oschkinat H (2004) The oxidized subunit
B8 from human complex I adopts a thioredoxin fold.
Structure 12, 1645-1654
Gaiser OJ, Ball LJ, Schmieder P, Leitner D, Strauss H,
Wahl M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
Structure, Backbone Dynamics, and Association
Behavior of the C-Terminal BRCT Domain from the Breast
Cancer-Associated Protein BRCA1. Biochemistry 43,
15983-15995
Strauss H, Hughes J, Schmieder P (2005) Heteronuclear
solution state NMR-studies of the chromophore in cyanobacterial
phytochrome Cph1. Biochemistry, 44, 8244-8250
Collaborations
K. Rueck-Braun, TU Berlin
P. Hildebrandt, TU Berlin
H. von Döhren, TU Berlin
T. Lamparter, FU Berlin
J. Hughes, Justus Liebig University Giessen
Structural Biology
19

STRUCTURAL BIOINFORMATICS
Group Leader: Dr. Gerd Krause
STRUCTURAL BIOINFORMATICS TO UNRAVEL
MOLECULAR SIGNALING MECHANISMS
The group is focused on genome and protein sequence
analysis by structural bioinformatics combined with comparative
modeling of protein-protein interaction to study
sequence- and structure- function relationships. The main
aim is rational discovery of molecular mechanisms for protein-protein
interactions and protein-ligand interaction to
deduce ideas for pharmacological intervention. All group
projects are characterized by close cooperation with
experimental partners to ensure a closed loop between
the theoretically derived model and experimental proof as
an important prerequisite for a successful application of
biocomputing methods. The group is concerned with two
topics to elucidate structure - function relationships of
protein-protein interactions and of G-protein coupled
receptors.
Protein-protein interactions
S. Müller, U. Wiedemann, G. Krause
Protein interaction networks are mediated very often by
non-catalytic protein domains like PDZ- and WW-domains.
Narrowing down potential interacting proteins from the
protein interaction network, elucidation of selective protein-protein
interaction patterns and subsequent identification
of complementary interaction sites at the levels of
epitopes, residues and atoms are the main aims.
• Interaction sites of junctional proteins
Junctional proteins connect and seal the contact sites in
between endothelial cells. The exact sites, structures and
molecular mechanisms of interaction between junction
organizing zona occludence protein 1 (ZO-1) and the tight
junction protein occludin or the adherens junction protein
α-catenin were unknown. Binding studies by surface plasmon
resonance spectroscopy and peptide mapping combined
with comparative modeling utilizing crystal structures
led for the first time to a molecular model revealing the
binding of both occludin and α-catenin to the same binding
site in ZO-1. Our data support a concept that ZO-1 successively
associates with α-catenin at the adherens
junction and occludin at the tight junction.
Strong spatial evidence indicates that the identified occludin
C-terminal coiled-coil domain dimerizes and interacts
as a helix bundle with the identified structural motifs in
ZO-1 (Müller et al. 2005). The selectivity of both protein-
protein interactions is defined by complementary shapes
and charges between the participating epitopes. In conclusion,
a common molecular mechanism of forming an
intermolecular helical bundle between the hinge region/
GuK domain of ZO-1 and α-catenin and occludin is identified
as a general molecular principle organizing the association
of ZO-1 at adherens and tight junctions (Schmidt et
al. 2004). The promising results of this joint project in collaboration
with the cell physiology group of I. Blasig, FMP,
were primarily due to the fact that the bioinformatic/
modeling- and the experimental studies as well have been
carried out by the same person (S.L. Müller). We are
currently extending the structure-function studies also to
other junctional proteins by combining structural information
(bioinformatics/comparative modeling) and biochemical-,
molecular biological studies.
• Prediction of protein – protein interaction network
partners
To better understand and to predict the interactions, predictive
interaction models (e.g. COMFA and profile hidden
Markov model recognition activities of WW domains)
were derived by U. Wiedemann from experimental screening
data. Taking the sequence (Otte et al. 2003) or the
structure (Schleinkofer et al. 2004) of experimentally undescribed
domains or ligands, these models allowed the
prediction of potential binding partners. Suggested optimal
peptide sequence for binding a subfamily of WWdomains
was successfully verified (Schleinkofer et al.
2004). Additionally, some models predicted the strength
(affinity) of the interaction (www.fmp-berlin.de/nmr/pdz).
Using this approach, ligands with higher affinity were
designed for representative domains of both families. In
particular, for the hAF6-PDZ, the hERBIN-PDZ and the
mSNA1-PDZ so-called “super-binding” peptides were
designed which indeed exhibited the highest affinity
achievable by combination of natural amino acids (Wiedemann
et al. 2004).
Molecular signal transduction mechanisms of
GPCR
G. Kleinau, J. Lättig, G. Krause
• Selective interaction patterns for G-protein subtypes
The molecular interactions at the interface between
G-protein-coupled receptors and the G-proteins is of high
functional importance, since there the decision takes
place about the different specific signaling pathways (via
the G-protein subtypes G αi, G αs, and G αq) into the cell. For
the thyroid stimulating hormone receptor (cooperation
with the University of Leipzig and NIH Bethesda, MD,
USA), we were able to delineate residues of the second
intracellular loop as selective recognition patterns for

C-terminal
tail
LRRX
binding site
Phe-Spine
G-protein subtypes by using molecular interaction models.
Site-directed mutagenesis confirmed M527 as a residue
selectively interacting with G αq (Neumann et al. 2005).
Our previous studies on peptides and other small molecules
(lipoamines) which directly interact with G-proteins
revealed potential interaction sites based on charge patterns.
Positive charges at ligands interact with negatively
charged residues at the G-protein site. Transferring these
complementary cognition patterns to the Endothelin receptors
led to identification of amino acids that might be
responsible for different signaling pathways of the receptor
subtypes. To provide support to this hypothesis, suggested
site-directed mutagenesis (J Lättig) is currently under
investigation by the group of Alexander Oksche (FU Berlin).
• Structural determinants of TSH- receptor activation at
the TSHR ectodomain:
Thyroid-stimulating hormone (TSH, thyrotropin) and the
TSH-Receptor (TSHR) are key proteins in the control of
thyroid function. TSH plays a critical role in ontogeny. As
a consequence, different mutations of TSHR and TSH
result in pathogenic diseases, for example thyroid toxic
adenomas, non-autoimmune and neonatal hyperthyroidism,
and hypothyroidism. Constitutively active TSH receptor
mutants (CAMs) are the molecular etiology of > 50% of
N-terminal
tail
N-terminal
Cysteine Box
LRRO
N-terminal
Cysteine Box-1
FIGURE 1
a: LRR structure of the Nogo-receptor ectodomain (pdb
entry 1OZN). A ‘Phe-spine’ (green aromatic rings) inside the
LRR is stabilizing the fold instead of the missing helices at
the concave outer face.
b: LRR domain: comparison of the previous model for TSHR
(pdb entry 1XUM, grey) based on ribonuclease inhibitor template
(pdb entry 2BNH) and the new generated homologous
LRR model for the TSHR (colored) based on the X-ray structure
of the hNogo-66-receptor ectodomain. The new template
provides the N-terminal flanking Cys-box1 as an integral
structural part of the LRR by contributing with an
additional parallel β-stand (LRR0 in red) to the convex
β-sheets of the proposed hormone-binding site.
hyperthyroidism in Germany. CAMs are thought to mimic
to some extent the active conformation of the wild type
(wt) receptor and to spontaneously adopt a structure able
to activate G-proteins. A concept for structure-function
relationships in processes of ligand-dependent and -independent
(constitutive) activity for TSHR would profoundly
modify the understanding of pathophysiology.
TSH belongs to the family of glycoprotein hormones. The
main differences of TSHR and the other glycoprotein hormone
receptors (GPHR) such as choriogonadotrophic/
luteinizing hormone receptor (CG/LHR) and follicle-stimulating
hormone receptor (FSHR) to other seven transmembrane-spanning
G-protein-coupling receptors (GPCRs) is
an even more complex activation mechanism, which
requires the binding of a large hormone towards a large
N-terminal ectodomain. The intramolecular mechanism of
the signal transduction to the serpentine domain upon hormone
binding at the ectodomain is not understood. The
elucidation of the intramolecular mechanisms of TSH
receptor signaling in the large TSHR ectodomain are a prerequisite
for establishing new perspectives for the treatment
of hyperthyroidism by inverse agonists or for the
treatment of thyroid cancer by small TSH receptor ligands.
Structural Biology
21

Cysteinebox-2
Cysteinebox-3
F405
D403
F 286
E404
N406
C284
C408
C283
S281
turn/loop
To identify determinants at the GPHR ectodomain that may
be involved in signal transduction, G. Kleinau first addressed
this issue by searching for homologous structural features
at the ectodomain based on high sequence similarity.
For the LRR motif within the large ectodomain, a new augmented
structural model based on the Nogo-66-receptor
ectodomain with 9+2 repeats was generated (Fig. 1a) that
resulted in a strongly enlarged radius at the hormone binding
site at the inner convex surface of the LRR-arch. The
structure is stabilized by an interior Phe-spine instead of
helices at the concave outer face based on previous templates
(Fig. 1b). Moreover, the sequence similarity of the
new template indicated that the N-terminal flanked
Cysteinbox-1 is also an integral structural part of the LRRarch
by contributing with an additional parallel (LRR0
(TSHR37-41)) anti-parallel ‚-strand to the convex ‚-sheet of
the hormone-binding region.
Furthermore, this model provides for the first time a structural
rationale for the previously observed participation of
residues from Cys-box1 in hormone binding, i.e. for three
residues at the additional repeat LRR0 of the FSHR 29-31 and
of TSHR 37-41. This LRR model of BHR was later on confirmed
by the X-ray structure of the LRR domain for FSHR
(Fan & Hendrichson, Nature 2005).
TM1
FIGURE 2
TSHR ectodomain: detail of Cys-box2 (red) and Cys-box-3
(yellow) interaction model adopted from IL8 and IL8RA complex
structure. Aromatic interaction of F 286 (C-b2) and F 405
(C-b3); Disulfide bridge between C 408 (C-b3) and C 283 (C-b2)
based on biochemical data (blue); S281-loop/turn conformation
adopted from Malonyl Coenzyme (cyan); The subsequently
predicted residues D 403, E 404, N 406 to be involved
in signal transduction at the interface between ectodomain
and serpentine domain indeed showed constitutive activities
upon mutations.
The subsequent structural extracellular component Cysbox2
is attached back-to-back to the 11th ‚-strand of the
LRR, very likely via a short turn/loop containing S 281. Extensive
systematic sequence similarity searches of fragmented
sequence portions of the ectodomain resulted in a
homologous model based on IL8-IL8RA fragment complex
(pdb entry 1ILQ (27) which links Cys-box 2 and Cys-box-3
together (Fig. 2).
Subsequently, the residues D 403EFNPC 408 of the C-b3 are
located particularly at prominent interface positions of the
ectodomain, most closely to the transmembrane domain.
The hydrophilic residues D 403, E 404 and N 406 are therefore
hypothesized to participate very likely in the intramolecular
signal transduction from the ectodomain towards the
serpentine domain (Kleinau et al, 2004).
Using our approach, we identified three new mutations
(D 403A, E 404K and N 406A) in the ectodomain of the TSHR that
are causing constitutive cAMP activity, and we suggest
that this motif is indeed important for the transduction of
the signal from the ectodomain to the transmembrane
domain. According to the high sequence conservation, the
results are of general relevance for the signal transduction
mechanism of other glycoprotein hormone receptors.

Group members
Sebastian L. Müller (Doctoral student)
Jens Lättig (Doctoral student)
Urs Wiedemann (Doctoral student)
Gunnar Kleinau (Doctoral student)
External funding
Deutsche Forschungsgemeinschaft
„Wechselwirkungen von Blut-Hirnschranken-Proteinen“
(Bl 208/6.2)
Ingolf Blasig, Gerd Krause
Deutsche Forschungsgemeinschaft
„Identifizierung eines rezeptorinternen stillen Transmitters
im TSH-Rezeptor“ (KR 1273)
Gerd Krause
Selected publications (FMP authors in bold)
Müller SL, Portwich M, Schmidt A, Utepbergenov DI,
Huber O, Blasig IE, Krause G (2005) The tight junction protein
occludin and the adherens junction protein α-catenin
share a common interaction mechanism with ZO-1. J Biol
Chem 280, 3747-3756
Schmidt A, Utepbergenov DI, Mueller SL, Beyermann M,
Schneider-Mergener J, Krause G, Blasig IE (2004) Occludin
binds to the SH3-hinge-GuK unit of zonula occludens
protein 1: potential mechanism of tight junction regulation.
Cell Mol Life Sci 61, 1354-1365
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Prot Sci 12, 491-500
Schleinkofer K, Wiedemann U, Otte L, Wang T, Krause G,
Oschkinat H, Wade RC (2004) Comparative Structural and
Energetic Analysis of WW Domain-peptide Interactions. J
Mol Biol 344, 865-881
Wiedemann U, Boisguerin P, Leben R, Leitner D, Krause
G, Mölling K, Volkmer-Engert R, Oschkinat H (2004) Quantification
of PDZ Domain Specificity, Prediction of Ligand
Affinity and Rational Design of Super-Binding Peptides. J
Mol Biol 343, 703-718
Neumann S, Krause G, Claus M, Paschke R (2005) Structural
Determinants in the Second Intracellular Loop for
G-Protein Selectivity of the Thyrotropin Receptor. Endocrinology
146, 477-485
Kleinau G, Jäschke H, Neumann S, Lättig J, Paschke R,
Krause G (2004) Identification of a novel epitope in the TSH
receptor ectodomain acting as intramolecular signalling
interface. J Biol Chem 279, 51590-51600
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V 2
receptors. J Biol Chem 279, 47254-47263
Collaborations
R. Paschke, TSH receptor,
University of Leipzig activation mechanism
S. Rothemund, IZKF Leipzig Peptide mapping
A. Oksche, FU Berlin V2 receptor
O. Huber, Charite Berlin CBF Interaction of junction
proteins
S. Offermanns, GPR109a receptor/
University of Heidelberg ligand interaction
S. Neumann, NIH, NIDDK TSH receptor
Bethesda, MD, USA
M. Gershengorn, NIH, NIDDK TSH receptor
Bethesda, MD, USA.
Structural Biology
23

MOLECULAR MODELLING
Group Leader: Dr. Ronald Kühne
COMPUTATIONAL TECHNIQUES FOR
LIGAND DESIGN AND PROTEIN MODELLING
The research of the Molecular Modelling/Ligand Design
Group is mainly focused on protein modeling, virtual
screening, and lead optimization in close collaboration
with experimental partners within academia as well as
industry. The special expertise of the group includes a
wide range of molecular modeling technologies, bioinformatics
tools, and computational chemistry methods.
On top of that, these methods are applied to support the
Screening Unit and the medicinal chemistry group within
the national ChemBioNet initiative. The focus in this field
is to analyze, design, and purchase a compound database
available for academic screening projects within the
framework of ChemBioNet. In addition to the already
ongoing modeling projects, the group has also started
research in several new fields during the last two years:
structure and function of antimicrobial peptides, high
throughput screening evaluation and virtual screening of
antigen-exchange-inducing agents targeting the major
histocompatibility complex (MHC) and inhibition of protein-protein
interaction in the IL6/IL-6R/gp130 complex. All
of these projects benefit from close collaborations with
groups within the FMP and the MDC as well as with industrial
partners.
Targeting G-protein-coupled receptors
G-protein-coupled receptor (GPCR) modeling and agonist
and antagonist design is one of the key competences of
the group. The applied computational tools range from
quantum chemically supported ligand design to high
throughput virtual screening of millions of compounds. The
GPCR's are modeled using the bovine rhodopsin x-ray
structure and refined using simulated annealing and molecular
dynamics in implicit as well as explicit lipid environment.
These models are exploited in virtual screening and
lead optimization using simulated annealing and pharmacophore
mapping.
The follicle-stimulating hormone receptor
(A. Schrey, R. Kühne)
We have constructed a model of the human follicle stimulating
hormone receptor (FSHR) in a water-vacuum-water
environment. The model is consistent with published mutational
data. A central feature of the model is a sodium ion
located between Asp408 transmembrane helix 2 (TM2),
Asp581 (TM6), and Asn618 and Asn622 (TM7). Activation
of the receptor seems to be associated with the translocation
of this ion and subsequent movements of TM 3, 6,
and 7. Further, we recognized a small molecule binding site
within the FSHR. Docking simulations supported by screening
results served as a basis for lead optimization processes.
Refined models of the human and rat FSHR derived
from molecular dynamics simulations in explicit lipid environment
have given closer insight into the interactions of
the residue Histidine615 (TM7), which is specific for the
human FSH receptor. We found that this residue located
towards TM6 is establishing a hydrogen bond to a backbone
carbonyl. In contrast to human FSHR, all known FSHR
of other species contain in this position a tyrosine residue.
In our simulations the tyrosine establishes hydrogen bonds
to residues in TM3. These results are able to explain the
known mutational data and other experimental facts. The
studies were sponsored by Schering AG.
Gonadotropin-Releasing Hormone Receptor
(A. Söderhäll, R. Kühne)
In cooperation with Zentaris AG (Frankfurt am Main), several
models of the human gonadotropin-releasing hormone
receptor (GnRH-R) have been derived. The models form a
series of successively improved generations of the receptor,
where each generation is based on the latest experimental
data. The data is based on site-directed mutagenesis,
activity modulation based on mutagenesis of the
native ligand as well as modulation of peptidomimetic
ligands. The latest generation of the receptor-ligand complex
was used to derive a pharmacophore pattern, which
was successfully used in the lead finding and optimization.
As a result of this successful cooperation with Zentaris,
two patents on new non-peptidic GnRH antagonists are
currently in process. One of them could be economically
utilized by the Forschungsverbund Berlin e.V.
Structure and function of antimicrobial peptides
(A. Söderhäll, F. Eisenmenger)
Antimicrobial peptides have recently emerged as a promising
new generation of antibiotics. This class of peptides
is generally believed to target the bacterial membrane
and destroy the chemical gradients over these, either
by building transmembrane pores or by destabilizing the
bilayer structure of the membranes. This mechanism of
action is believed to make the peptides more stable
against the evolutionary pressure from bacteria developing
antibiotics resistance. By exploiting the in-house
competence on peptide chemistry in Michael Bienert’s
group, the antimicrobial peptide cyclo-(RRWWRF) was
selected for NMR structure determination by Christian
Appelt in the group of Peter Schmieder. Using the NMR

–10
–5
0
5
10
structure we have simulated realistic membranes at various
peptide-to-lipid ratios. From these molecular dynamics
simulations we have concluded that the peptide disturbs
the membrane so that its function as a hydrophobic barrier
becomes less effective, and may increase the permeability
of ions as well as water. Interestingly, the perturbation
of the hydrophobic barrier takes place without the
formation of explicit pores, as was often suggested for this
kind of substances. We have elucidated in detail how this
compound interacts with the membrane lipids and what
impact this has on the membrane structure /see Fig.1/.
High throughput and virtual screening targeting
the major histocompatibility complex
(A. Söderhäll, R. Kühne)
The group of Olaf Rötzschke at MDC has screened the
FMP database for antigen-exchange inducing agents targeting
the MHC. The data from the high throughput screen
of the 20,000 compounds has been structured and analyzed
in detail. The analysis was then used in the construction
of a prediction model based on the quantitative structure-activity
relationships (QSAR) of a hit subfamily from
the high throughput screening campaign. Based on this
derived model, an extended virtual screening of three
10
5
0
–5
–10
FIGURE 1
The structure of the antimicrobial peptide cyclo-(RRWWRF)
was determined by NMR and exploited in a series of molecular
dynamics simulations in the presence of explicit lipids.
The figure shows the average structure of cyclo-(RRWWRF)
from the simulation and a surface representation of the
membrane. The amphipatic character of the peptide creates
a void in the membrane surface that contributes to the
increased permeability of water and most likely also ions
through the membrane.
external databases containing a total of more than one million
compounds was carried out. This massive data set
was narrowed down to a list of 5589 ranked compounds.
A dozen of these were hand-picked and experimentally
tested. Five out of these twelve were found to be active,
proving that the hit enrichment of the focused library was
of major help in the second, refined screening round by O.
Rötzschkes group. Within the framework of this project,
dockings of the lead compounds were carried out in order
to explain the mechanisms behind the antigen exchange.
The docking approach was supported by site-directed
mutagenesis. Using these dockings together with experimental
data, we are able to propose a possible mechanism
of action of antigen exchange.
NMR structure calculations
(C. Brockmann, R. Kühne)
An important part of the scientific activities is related to
structure elucidation of protein domains by NMR. The
main focus in this field is on NMR structure calculations
and methods to refine NMR derived protein structures.
NADH ubiquinone oxidoreductase (complex I) is the last
major complex of the respiratory chain for which there is
no detailed atomic structure. Since most proteins of this
Structural Biology
25

A B
R [cm]
highly modular complex are rather small, its components
make suitable targets for structural studies by NMR. In this
project the structure of subunit B8 (CI-B8) was investigated
by NMR. The solution structure of the oxidized subunit
shows a thioredoxin fold with remarkable similarities to
thioredoxin-like Fe2S2-ferredoxins (Fig. 2A). A structural
comparison shows high similarities of surface properties
and possibly active residues. Since CI-B8 shows some
similarities to thioredoxins, and since the mammalian
homologues may form a disulfide bridge in a similar position,
we determined the redox potential of oxidized CI-B8
to see if it fits into the range covered by active site disulfides
in thioredoxin-related proteins (Fig. 2B). The redoxpotential
of the disulfide bond of CI-B8 compares well to
that of the catalytically active disulfides in other thioredoxin-like
proteins. This, in addition to the fact that CI-B8
stems from a complex that is involved in a redox-dependent
reaction, leads to the idea that CI-B8 could assist in
redox-associated processes or electron transfer. This
example of CI-B8 shows that structural studies of the individual
components of complexes can spark hypotheses
about the function of the component within the complex.
1,0
0,8
0,6
0,4
0,2
0,0
1E-4 1E-3 0,01 0,1 1 10 100 1000
[GSH] 2 / [GSSG]
FIGURE 2
Structure and redox-potential determination of CI-B8. A: ribbon-representation of the structure of human CI-B8. Alpha-helices are depicted in
green, the beta-sheet in blue. The position of the disulfide-bond is indicated in yellow. B: Determination of the redox potential. The change in
Trp-fluorescence upon reduction is plotted against the relative concentrations of the redox-buffer system. From a Keq of 0.486 a redox potential
of –250 mV was calculated.
Group members
Dr. Anna Schrey
Dr. Arvid Söderhäll
Dr. Jörg Wichard*
Dr. Frank Eisenmenger (Unix system administration)
Christoph Brockmann (Doctoral student)
Stefan Hübel (Unix system administration support, database
management)
External funding
Schering AG
Transmembranrezeptoren (Kooperationsvertrag)
Zentaris AG
(Kooperationsvertrag)
Conaris AG
Selected publications (FMP authors in bold)
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kühne R, Oschkinat H (2004) The oxidized subunit
B8 from human complex I adopts a thioredoxin fold.
Structure (Cam) 12, 1645-1654
* part of period reported

Gaiser O, Ball LJ, Schmieder P, Leitner D, Strauss H, Wahl
M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
structure, backbone dynamics, and association behavior
of the C-terminal BRCT domain from the breast cancer
associated protein BRCA1. Biochemistry 43, 15983-15995
Pope BJ, Zierler-Gould KM, Kühne R, Weeds AG, Ball LJ
(2004) Solution structure of human cofilin: actin binding,
pH sensitivity, and relationship to actin-depolymerizing
factor. J Biol Chem 279, 4840-4848
Brockmann C, Leitner D, Labudde D, Diehl A, Sievert V,
Bussow K, Kühne R, Oschkinat H (2004) The solution structure
of the SODD BAG domain reveals additional electrostatic
interactions in the HSP70 complexes of SODD subfamily
BAG domains. FEBS Lett 558, 101-106
Freund C, Kühne R, Park S, Thiemke K, Reinherz EL, Wagner
G (2003) Structural investigations of a GYF domain
covalently linked to a proline-rich peptide. J Biomol NMR
27, 143-149
Zimmermann J, Kühne R, Volkmer-Engert R, Jarchau T,
Walter U, Oschkinat H, Ball LJ (2003) Design of N-substituted
peptomer ligands for EVH1 domains. J Biol Chem
278, 36810-36818
Karges B, Karges W, Mine M, Ludwig L, Kühne R, Milgrom
E, de Roux N (2003) Mutation Ala(171)Thr stabilizes the
gonadotropin-releasing hormone receptor in its inactive
conformation, causing familial hypogonadotropic hypogonadism.
J Clin Endocrinol Metab 88, 1873-1879
Collaborations
Dr. W. Karges, University of Ulm
Prof. Schmalz, University of Cologne
Dr. O. Roetzschke, MDC
Structural Biology
27

SOLID STATE NMR
Group Leader: Prof. Bernd Reif
SOLID-STATE NMR SPECTROSCOPY:
PROTEIN AGGREGATION AND MEMBRANE
PROTEINS
We use Nuclear Magnetic Resonance (NMR) in order to
characterize biomolecular systems which are situated at
the interface between solution and solid. In this area,
membrane proteins and amyloidogenic peptides and proteins
are the most interesting “target molecules”. By
nature, structural information of these systems is difficult
to obtain by means of X-ray crystallography or standard
solution-state NMR methods. We want to address these
systems by application of a combination of modern solution
state and solid state NMR methods. This requires
development of especially adapted NMR techniques. So
far, about 20 proteins are known for which a correlation
between aggregation and disease is established. The
most prominent examples are Alzheimer's disease, the
prion diseases (BSE, CfJ), and Huntington disease. However,
little is known about the mechanism which leads to
aggregation, as well as on the structure of the amyloid
fibrils. We would like to gain more insight into the structure
of oligomeric intermediate states which are associated
with protofibril formation. In addition, we are interested in
characterizing dynamic chemical exchange processes
between the soluble and aggregated state of the respective
proteins.
Yeast Prions: Sup35 and Hsp104
Characterization of interactions between the molecular
chaperone Hsp104 and the prion protein Sup35 in yeast by
NMR spectroscopy Sup35 is a subdomain of the translation-termination
complex in S. cerevisiae. Under specific
conditions, Sup35 can form protein aggregates that can
induce a conformational change in other Sup35 proteins
and are inherited after cell division. Hsp104 is one of the
most important proteins for the thermostability in yeast and
is, together with two other proteins, Hsp70/Ssa1 and
Hsp40/Ydj1, involved in a chaperon complex that can dissolve
protein aggregates that are produced as a heat
response. Solution-state NMR methods are employed to
characterize the interactions between the aggregate and
the chaperone. We found that Hsp104 is able to disaggregate
Sup35-derived peptides in vitro, and that especially
low oligomeric complexes of Sup35 interact with Hsp104
(Narayanan et al., 2003). In this study, we were able to analyze
conformational changes of Sup35 induced by Hsp104
by NMR in real time.
Structural characterization of ligands targeted
against beta-amyloid fibrils (Aß)
Alzheimer's disease (AD) is the most common form of agerelated
neurodegenerative disorder, and is related to
deposition of Aβ peptides in the brains of AD patients. Aβ
is formed by processing of APP, the Amyloid Precursor
Protein, a membrane protein of yet unknown function. We
use the fact that amyloid fibrils orient spontaneously in the
magnetic field to determine the structure of peptide inhibitors
that bind to Aβ aggregates (Chen & Reif, 2004). The
alignment is based on the magnetic anisotropy of the peptide
bond. A net alignment of amyloid fibrils is induced due
to the arrangement of hydrogen bonds in parallel to the
fibril axis. We expect that this approach is useful for the
design of diagnostic or therapeutic ligands.
Structural characterization of acid denatured
PI3K fibrils
Chris Dobson and co-workers demonstrated that not only
proteins which are related to a disease can form highly
structured aggregates, but almost any other soluble protein
under certain solution conditions. One example is the
SH3 domain of the phosphatidyl-inositol-3-kinase (PI3K)
which forms fibrils under acidic pH. Solid-state NMR experiments
revealed that His25 of PI3K-SH3 is involved in tertiary
contacts which stabilize the fibril structure (Ventura
et al., 2004). Mutational studies (H25K) confirmed this
hypothesis. Most importantly, it could be shown that artificial
neurotoxicity is abolished in the mutated protein
using a MTT reduction assay. We use PI3K-SH3 as a paradigm
to study intermediate states as well as the aggregated
state and try to obtain a better understanding of the
mechanism of protein aggregation.
Development of MAS solid-state NMR techniques
Solid-state techniques have found great attention in the
last few years since it has turned out to be possible to
determine the structure of uniformly labeled (crystalline)
peptides and proteins (Rienstra et al. 2002). However,
solid-state NMR experiments are intrinsically insensitive
due to detection of the heteronucleus (carbon or nitrogen).
We address the question of sensitivity by employing proton
detection in combination with deuteration, which chemically
eliminates the strong proton-proton dipolar couplings.
This approach allows to detect proteins with high
sensitivity (Chevelkov et al. 2003). In addition, long range
proton-proton interactions are accessible, which are
important to determine the three-dimensional fold of a protein
in the solid state (Reif et al. 2003). Furthermore, we
explore deuterium as a source for dynamic information in
the solid state.

Sup35
Hsp104 +
Sup35
Monomeric
Sup35
Group members
Dr. Alex Chernogolov
Dr. Maggy Hologne
Zhongjing Chen (Doctoral student)
Veniamin Chevelkov (Doctoral student)
Muralidhar Dasari (Doctoral student)
Saravanakumar Narayanan (Doctoral student)
Uwe Fink (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Entwicklung NMR spektroskopischer Methoden zwischen
Flüssigkeit und Festkörper. Strukturuntersuchungen
an orientierten Biomakromolekülen“ (Re1435/2)
Bernd Reif
Deutsche Forschungsgemeinschaft
„Strukturelle Charakterisierung des Multidrug-Transporters
EmrE mittels MAS Festkörper-NMR-Spektroskopie“
(Re1435/3)
Bernd Reif
Higher
Oligomeric
States of
Sup35
Intermediate 1
Intermediate 2
FIGURE 1
Hsp104 interacts preferably with low oligomeric Sup35.
Interactions between Sup35 and Hsp104 are characterized
by STD NMR experiments. Sup35 oligomeric states are
determined using DOSY experiments.
Deutsche Forschungsgemeinschaft
„Biochemische und Strukturuntersuchungen an Sup35p
im Komplex mit Hsp104, Hsp40 und Hsp70“ Teilprojekt A3
im Sonderforschungsbereich 594 „Molekulare Maschinen“
Bernd Reif
Selected publications (FMP authors in bold)
Chen Z, Reif B (2004) Measurement of residual dipolar
couplings in peptidic inhibitors weakly aligned by transient
binding to peptide amyloid fibrils. J Biomol NMR 29, 525-
530
Chevelkov V, Rossum BJv, Castellani F, Rehbein K, Diehl
A, Hohwy M, Steuernagel S, Engelke F, Oschkinat H, Reif
B (2003) 1H detection in MAS solid state NMR spectroscopy
employing pulsed field gradients for residual solvent
suppression. J Am Chem Soc 125, 7788-7789
Narayanan S, Bösl B, Walter S, Reif B (2003) Importance of
low oligomeric weight species for prion propagation in the
yeast prion system Sup35/Hsp104. Proc Natl Acad Sci USA
100, 9286-9291
Structural Biology
29

Reif B, van Rossum BJ, Castellani F, Rehbein K, Diehl A,
Oschkinat H (2003) Determination of 1H 1H distances in a
uniformly 2H,15N labeled SH3 domain by MAS solid state
NMR spectroscopy. J Am Chem Soc 125, 1488-1489
Rienstra CM, Tucker-Kellogg L, Jaroniec CP, Hohwy M,
Reif B, McMahon MT, Tidor B, Lozano-Pérez T, Griffin RG
(2002) De Novo Determination of Peptide Structure with
Solid-State MAS NMR Spectroscopy. Proc Natl Acad Sci
USA 99, 10260-10265
Ventura S, Zurdo J, Narayanan S, Parreño M, Mangues R,
Reif B, Chiti F, Giannoni E, Dobson CM, Aviles FX, Serrano
L (2004) Short amino acid stretches can mediate amyloid
formation in globular proteins: Thr Src homology 3 (SH3)
case. Proc Natl Acad Sci USA 101, 7258-7263
Collaborations
Fibril Axis
Magnetic Field
Dr. W. Boelens, University of Nijmegen, The Netherlands
Prof. J. Buchner, TU Munich
Dr. G. Gast, University of Potsdam
Prof. Dr. U. Heinemann, MDC Berlin
Prof. Dr. W. W. de Jong, University of Nijmegen, The
Netherlands
Prof. G. Multhaup, Free University Berlin
FIGURE 2
Structure of a peptide inhibitor bound to an Aß fibril using
restrained docking. The spontaneous alignment of fibrils in
the magnetic field induces an ordering of the ligand peptide.
Measurement of transfer residual dipolar couplings
(trRDCs) yield the relative orientation of the peptide with
respect to the fibril axis.
Dr. M. Sattler, EMBL Heidelberg
Prof. Dr. S. Schuldiner, Hebrew University Jerusalem,
Israel
Dr. L. Serrano, EMBL Heidelberg
Dr. S. Ventura, Universitat Autonoma de Barcelona,
Spanien.
Dr. S. Walter, TU Munich

PROTEIN ENGINEERING
Group Leader: PD Dr. Christian Freund
PROTEIN ADAPTER DOMAINS INVOLVED IN
T CELL SIGNALING
Our group is interested in the molecular interactions that
govern the assembly of protein complexes. The focus is
on adapter domains that mediate protein-protein interactions
in immune cells, especially T cells. The intracellular
response in T cells has to cope with the highly adaptive T
cell receptor engagement process. Subtle changes in the
kinetics and affinity of the TCR-MHC-peptide interaction
are able to evoke dramatically different immune responses.
The fine-tuning of the intracellular processes is
dependent on the membrane organization, the molecular
patterning at the inner membrane and cytoskeletal rearrangements
that take place upon stimulation. Cytoplasmic
adapter domains that guide the spatial distribution of
proteins are present in many lymphoid-specific as well as
general signaling molecules. They typically recognize peptide
sequences that are present in the cytoplasmic
domains of transmembrane receptors or within solvent
exposed regions of intracellular proteins. Deciphering this
sequence recognition code and determining the regulatory
mechanisms of such adapter domain mediated interactions
is the major topic in our group. Therefore we employ
NMR spectroscopy, protein biochemistry, fluorescence
microscopy and screening of biomolecular libraries as
research tools. Protein Engineering approaches in our
laboratory implement random and structure-based mutagenesis
as strategies to derive biologically relevant information.
Based on the determination of the three-dimensional
structure of protein domains, we want to understand
recognition at a molecular level, while biochemical studies
are aimed at putting this knowledge in the context of
cellular signaling.
CD2BP2
Michael Kofler, Matthias Heinze, Katharina Thiemke, and
Christian Freund
In addition to its binding capacity for the CD2 cytoplasmic
domain, we have identified novel in vitro binding partners
for the CD2BP2 protein by means of peptide spot analysis
and subsequent NMR analysis (Kofler et al. 2004, J. Biol.
Chem. 279, 28292-28297). Several spliceosomal proteins
contain recognition sequences that match the requirements
for binding to the GYF domain of CD2BP2. Especially
the core splicing protein SmB/B’ contains several
binding motifs for CD2BP2-GYF, and we have determined
the epitope for the interaction. Pulldown experiments
confirm a specific interaction between the CD2BP2 and
SmB/B’ proteins, and fluorescence microscopy of live
cells shows the colocalization of the two proteins in the
nucleus of Jurkat and HeLa cells (Fig. 1). Based on our in
vitro results we are now testing additional proteins for
their potential to recognize the CD2BP2 protein in vivo.
GYF domains
Michael Kofler, Katrin Motzny, and Christian Freund
The NMR structure of this domain defines a new fold that
is proposed to be present in many eukaryotic proteins. A
conserved set of hydrophobic and aromatic amino acids
defines the binding site for the proline-rich ligand. The
structure of the GYF domain in complex with the
SHRPPPPGHRV peptide derived from CD2 reveals that the
prolines of the ligand form a polyproline type II helix that
contact the hydrophobic hot spot of the domain (Freund et
al. 2002, EMBO J 21, 5985-5995). The glycine residue
promotes a kink in the peptide backbone conformation,
thereby allowing the proline helix to be optimally placed
within the GYF domain binding pocket. Peptide substitution
analysis shows the importance of the proline-prolineglycine
motif for recognition by the GYF domain of CD2BP2.
We are currently investigating several other GYF domains
from various eukaryotic species by phage display. This
will allow us to compare the ligand sequence space of
these GYF domains, and structural studies are underway
that complement the peptide binding results. Mapping the
sequence space of GYF domains is anticipated to create a
valuable tool for the identification of in vivo interaction
partners. It will also allow us to better understand the
sequence requirements for individual subclasses of GYF
domains and set the stage for the comparison of sequence
recognition between GYF, SH3-, WW-domains.
Cyclophilin
Kirill Piotukh and Christian Freund
Cyclophilins contain an intrinsic peptidyl-prolyl-cis-trans
isomerase activity (Fischer et al. 1989, Nature 337, 476-
478), but have also been characterized as binding modules
within protein complexes (for a review see Ivery 2000, Med
Res Rev 20, 452-484). We are currently investigating the
potential of cyclophilin to act on and bind to proline-rich
sequences that are also ligands for proline-rich sequence
recognition domains. Conceptually it is believed that
cyclophilins recognize peptide conformation rather than
being highly sequence-specific. We will use in vitro
methods for the identification of peptide sequences that
can be recognized by cyclophilins. These peptides are
further investigated by NMR spectroscopy and other biophysical
methods to gain an understanding of the confor-
Structural Biology
31

A
C
B
ADAP hSH3 FYN SH3
mational restraints imposed on the peptides upon complex
formation.
ADAP
Katja Heuer, Marc Sylvester, Jürgen Zimmermann,
Katharina Thiemke, and Christian Freund
ADAP (adhesion- and degranulation- promoting adapter
protein) was identified as an adapter protein that upon
association with the TCR complex becomes tyrosine phosphorylated
and thereby creates binding sites for the SH2
domains of the Fyn kinase and of the SLP-76 protein (da
Silva et al. 1997, Proc Natl Acad Sci 94, 7493-7498; Musci
et al. 1997, J Biol Chem 272, 11674-11677). ADAP has also
been shown to colocalize with F actin and cytoskeletal
proteins such as VASP and WASP in activated Jurkat T
cells. A similar complex has been found enriched in the
phagocytic cups of activated macrophages (Coppolino et
al. 2001, J Cell Sci 114, 4307-4318; Krause et al. 2000, J Cell
Biol 149, 181-194). Furthermore, ADAP-deficient mice show
a decrease in LFA-1 dependent adhesion of stimulated
peripheral T cells (Griffiths et al. 2001, Science 293, 2260-
2263; Peterson et al., 2001, Science 293, 2263-2265) and
therefore ADAP was concluded to be an important regulator
for inside-out signaling in T cells and other hemato-
D
FIGURE 1
Comparison of the structure
of the ADAP hSH3 domain
with a “classical” SH3 domain.
The a-helix of ADAP
hSH3 contacts the SH3 fold
and several residues that are
important for binding the proline-rich
ligand are mutated in
ADAP hSH3 (compare A and
B, in B the peptide ligand is
shown in green). These features
lead to an altered surface
topology and the inability of
ADAP hSH3 to bind to prolinerich
sequences (compare C
and D).
poietic cells. From a molecular perspective, ADAP represents
a new member of the increasingly large family of
non-enzymatic, hematopoietic scaffolding proteins. In
addition to tyrosine-phosphorylation sites, ADAP contains
a number of N-terminal proline-rich motifs that mediate
the interaction with the SH3 domain of SKAP55 (Liu et al.
1998, Proc Natl Acad Sci 95, 8779-8784; Marie-Cardine et
al. 1998, J Biol Chem 273, 25789-25795). ADAP itself contains
two regions of homology to SH3 domains and we
have recently solved the structure of the ADAP C-terminal
domain by NMR spectroscopy (Heuer et al. 2004, Structure
12, 603-610). The structure reveals an extended SH3
domain fold, where an N-terminal α-helix contacts the SH3
scaffold, thereby creating a composite surface that cannot
bind to proline-rich sequences (Figure 2). We are currently
investigating the binding properties of this domain,
since we believe this will contribute to elucidation of
ADAP’s function within immune cell signaling.

Group members
Dr. Katja Heuer
Dr. Kirill Piotukh
Dr. Jürgen Zimmermann*
Michael Kofler (Doctoral student)
Matthias Heinze (Doctoral student)
Marc Sylvester (Doctoral student)*
Katharina Thiemke (Technical assistance)*
Ulrike Schneeweiß (Technical assistance)*
Uta Ben-Slimane (Technical assistance)** *
Katrin Motzny (Technical assistance)**
External funding
Bundesministerium für Bildung und Forschung
„Struktur-Funktionsbeziehung wichtiger T-Zell-Proteine
und Design von Agonisten und Antagonisten der T-Zell
vermittelten Immunantwort“ (Bio-Future 0311879)
Sieger im Nachwuchsgruppenwettbewerb Bio-Future
C. Freund
* part of period reported
** part-time
FIGURE 2
Binding site of the CD2BP2-GYF domain in complex with a
CD2 peptide. Binding site residues of the GYF domain are
shown in magenta while the peptide is displayed in yellow.
The surface of the domain is rendered transparent.
Volkswagen Foundation
“Biological Function of Adaptor Domains Controlled by The
Activity of Peptidyl-Prolyl cis/trans Isomerases” (I/77 955)
C. Freund
Deutsche Forschungsgemeinschaft
„Struktur-Funktionsbeziehung der GYF-Domäne“ (FR
1325/2-1)
C. Freund
Selected publications (FMP authors in bold)
Heuer K, Arbuzova A, Strauss H, Kofler M, Freund C (2005)
The helically extended SH3 domain of the T cell adaptor
protein ADAP is a novel lipid interaction domain. J Mol
Biol, in press
Heuer K, Kofler M, Langdon M, Thiemke K, Freund C (2004)
Structure of a helically extended SH3 domain of the T cell
adapter protein ADAP. Structure 12, 603-610
Kofler M, Heuer K, Zech T, Thiemke K, Freund C (2004)
Recognition sequences for the GYF domain reveal a possible
spliceosomal function of CD2BP2. J Biol Chem 279,
28292-28297
Structural Biology
33

Kofler M, Motzny K, Freund C (2005) GYF domain proteomics
reveals interaction sites in known and novel target
proteins. Mol Cell Proteomics (in press)
Freund C (2004) The GYF domain. In: Modular Protein
Domains, Wiley-VCH GmbH, Germany
Freund C, Kühne R, Park S, Thiemke K, Reinherz EL, Wagner
G (2003) Structural investigations of a GYF domain
covalently linked to a proline-rich peptide. J Biomol NMR
27, 143-149
Freund C, Kühne R, Yang H, Park S, Reinherz EL, Wagner G
(2002) Dynamic interaction of CD2 with the GYF and the
SH3 domain of compartmentalized effector molecules.
EMBO J 21, 5985-5995
Collaborations
Ellis Reinherz (Dana-Farber Cancer Institute, Boston, MA,
USA)
Volkhard Helms (University of Saarland, Saarbrücken)
Ingo Schmitz (University of Düsseldorf)
Richard Kroczek (Robert Koch Institute, Berlin)

CELLULAR SIGNALLING/MOLECULAR GENETICS

SECTION CELLULAR SIGNALLING/
MOLECULAR GENETICS
Prof. Ivan Horak
Department Head: Molecular Genetics
(Secretary: Alexandra Kiesling)
Prof. Walter Rosenthal
Department Head: Cellular Signalling
(Secretary: Heidemarie Petschick)
Research in the FMP section “Cellular Signalling / Molecular
Genetics” covers some of the major aspects of eucaryotic
signal transduction. Increasingly, the development
of pharmacological strategies to interfere with cellular
signal transduction pathways under study has become a
major point of emphasis.
The Protein Trafficking group is interested in the early stages
of the intracellular trafficking of G-protein-coupled
receptors (GPCRs). The intracellular transport of membrane
proteins starts with their insertion into the membrane
of the endoplasmic reticulum (ER). During ER insertion, the
INTRODUCTION
BEREICH SIGNALTRANSDUKTION/
MOLEKULARE GENETIK
Prof. Ivan Horak
Abteilungsleiter: Molekulare Genetik
(Sekretariat: Alexandra Kiesling)
Prof. Walter Rosenthal
Abteilungsleiter: Signaltransduktion
(Sekretariat: Heidemarie Petschick)
Die Forschung im FMP-Bereich „Signaltransduktion/
Molekulare Genetik“ befasst sich mit wichtigen Aspekten
der eukaryotischen Signaltransduktion. Zunehmend liegt
das Augenmerk auf der Entwicklung pharmakologischer
Strategien zur Interferenz mit den untersuchten Signaltransduktionswegen.
Die Arbeitsgruppe Protein Trafficking interessiert sich für
die frühen Stadien des intrazellulären Transports G-Protein-gekoppelter
Rezeptoren (GPCRs). Der intrazelluläre
Transport von Membranproteinen beginnt mit ihrer Insertion
in die Membranen des endoplasmatischen Retikulums
(ER). Unter der Insertion werden die Proteine gefaltet.
Dabei wird die korrekte Faltung durch ein Qualitätskontrollsystem
überwacht. Nur korrekt gefaltete Proteine tre-
proteins are folded, and correct folding is monitored by a
quality control system (QCS). Only correctly folded proteins
are allowed to enter the vesicular transport via the
ER/Golgi intermediate compartment (ERGIC) and the Golgi
apparatus to the plasma membrane. The study of the early
stages of the intracellular transport of GPCRs is also of
clinical significance. Naturally occuring receptor mutations
frequently lead to misfolded proteins unable to pass
the QCS and consequently to inherited diseases.
The aim of the Anchored Signalling group is to identify further
proteins involved in the redistribution of the protein
AQP2 that belongs to the family of water channels. It is critically
involved in the Arginine-vasopressin mediated
water reabsorption in the kidney. Arginine-vasopressin
binds to the vasopressin V2 receptor located on the surface
of renal principal cells thereby triggering the redistribution
of AQP2 from intracellular vesicles into the urinefacing
plasma membrane. The insertion of AQP2 into the
plasma membrane introduces water-selective pores and
facilitates water entry into the cells. The studies in this
group may lead to a detailed understanding of the under-
ten in den vesikulären Transport über das „ER-cis-Golgi
intermediate Compartment“ (ERGIC) und den Golgi-Apparat
zur Plasmamembran ein. Das Studium der frühen Stadien
des intrazellulären Transports von GPCRs ist auch von
klinischer Bedeutung. Natürlich auftretende Rezeptormutationen
führen häufig zu fehlgefalteten Proteinen, die
das Qualitätskontrollsystem nicht passieren können. Auf
diese Weise manifestieren sich Erbkrankheiten.
Das Ziel der Arbeitsgruppe Anchored Signalling ist es, Proteine
zu identifzieren, die an der Umverteilung des Proteins
AQP2 mitwirken. AQP2 gehört zur Familie der Wasserkanäle
und ist an der durch Arginin-Vasopressin-vermittelten
Wasserreabsorption in der Niere beteiligt. Arginin-
Vasopressin bindet an den Vasopressinrezeptor V2, der
sich auf der Oberfläche der Prinzipalzellen der Niere befindet.
Auf diese Weise wird die Umverteilung AQP2 aus
intrazellulären Vesikeln in die harnseitige Plasmamembran
ausgelöst. Durch das Eintreten von AQP2 in die Plasmamembran
wird diese mit wasserselektiven Poren ausgestattet,
so dass Wasser in die Zellen aufgenommen werden
kann. Die Untersuchungen in der Arbeitsgruppe sollen
zu einem detaillierten Verständnis des molekularen
Mechanismus der Wasserreabsorption führen und letztlich
eine Behandlung bestimmter Fälle des Diabetes insi-

lying mechanism, and eventually to a treatment of certain
cases of diabetes insipidus. Recently, interference of protein-protein
interactions by the means of chemical compounds
has become a major aspect.
The main field of the Biophysics Group is research in membrane
transport. Basic research projects aim to explore the
molecular mechanisms of water and proton movement.
The activities in applied biophysics are devoted to the control
of membrane permeability by (bio)polymers and to photodynamic
reactions of membrane components. Peter Pohl
has been appointed full professor at the University of Linz
in Austria and left the FMP at the end of 2004.
The research activity of the Department of Molecular
Genetics concerns the molecular mechanisms which control
development and function of cells in the blood and
immune system. Elucidation of these processes is an ultimate
requirement not only for understanding their pathological
alterations but also for the development of rational
diagnostic and therapeutic procedures. We have been
using targeted mutagenesis to analyze genes thought to
have important regulatory functions in the blood and
pidus möglich machen. Aktuell hat sich insbesondere die
mittels chemischer Verbindungen auslösbare Interferenz
mit den Protein-Protein-Interaktionen zu einem wesentlichen
Aspekt der Forschungstätigkeit entwickelt.
Der Hauptforschungsgegenstand der Arbeitsgruppe Biophysik
ist der Transport an Membranen. Forschungsprojekte
zielten auf die molekularen Mechanismen der Wasser-
und Protonenbewegung. Die Aktivitäten auf dem
Gebiet der angewandten Biophysik sind darauf gerichtet,
die Durchlässigkeit der Membran für (Bio-)Polymere und
die photodynamischen Reaktionen von Membrankomponenten
zu erforschen. Der Leiter der Gruppe, Peter Pohl,
folgte Ende 2004 einem Ruf auf einen Lehrstuhl an der Universität
Linz in Österreich.
Die Abteilung Molekulare Genetik befasst sich mit den
molekularen Mechanismen, die die Entwicklung und die
Funktion von Zellen des Blutes und des Immunsystems
kontrollieren. Die Erforschung dieser Prozesse ist eine
unabdingbare Voraussetzung nicht nur für das Verständnis
ihrer pathologischen Veränderungen sondern auch für die
Entwicklung rationaler diagnostischer und therapeutischer
Verfahren. Die Wissenschaftler nutzen zielgerichtete
Mutagenese, um Gene zu analysieren, von denen anzunehmen
ist, dass sie wichtige regulatorische Funktionen
immune system. This technology is used to generate
mouse mutants (“knock-out mice“) that carry an experimentally
designed gene defect. Thus it is possible to analyze
the consequences of a specific gene defect in the
context of the whole organism. The main interest is focused
on cytokine receptor signaling, in particular interferons.
These cytokines are used for treatment of human
diseases, despite their unknown functional mechanisms.
The highly pleiotropic effects of these cytokines are often
therapeutically disadvantageous. Therefore, an exact
knowledge of signaling pathways and interacting molecules
which are regulated by these cytokines could lead to
the development of more precisely targeted therapeutic
interventions.
Cytokine receptor signaling involves STATs (Signal transducers
and activators of transcription). Frequently, abnormal
activity of certain STAT family members, particularly
Stat3 and Stat5, is associated with a wide variety of human
malignancies, including haematological, head and neck,
breast and prostate cancers. The EMBO research group
of Uwe Vinkemeier is investigating the relationship bet-
im Blut und Immunsystem haben. Mit Hilfe dieser Methode
werden experimentell Mäuse mit einem Gendefekt
erzeugt (Knockout-Mäuse). Auf diese Weise ist es möglich,
die Auswirkungen eines spezifischen Gendefekts im
Kontext des Gesamtorganismus zu analysieren. Das
Hauptinteresse der Abteilung gilt dem Zytokinrezeptor-
Signaling, insbesondere aber den Interferonen. Diese
Zytokine werden zur Behandlung von Erkrankungen des
Menschen eingesetzt, man kennt jedoch ihre Wirkmechanismen
nicht genau. Der stark pleiotrope Effekt der Interferone
ist meist von Nachteil für die Therapie. Deshalb
können eine genaue Kenntnis der Signaltransduktionswege
und der Molekülinteraktionen, die durch die Zytokine
reguliert werden, zur Entwicklung präziserer therapeutischer
Interventionen führen.
Das Zytokinrezeptor-Signaling verläuft über STATs (Signal
transducers and activators of transcription). Eine abnormale
Aktivität bestimmter Mitglieder der STAT-Familie, insbesondere
von Stat3 und Stat5, steht im Zusammenhang
mit einer Vielzahl bösartiger Krebserkrankungen. Die
EMBO-Forschungsgruppe von Uwe Vinkemeier untersucht
die subzelluläre Lokalisation der STATs und die damit
zusammenhängenden Konsequenzen für die Gen Induktion.
Eine der Herausforderungen ist es, herauszufinden,
37 Cellular Signalling/ Molecular Genetics

ween the subcellular localization of STATs and the resulting
consequences on gene induction. One of the challenges
is to determine how target gene access is regulated
and how this might influence the execution of transcriptional
programs. The goal is a complete description of the
mechanisms underlying STAT subcellular trafficking by
focusing on the biochemical basis of nuclear accumulation
as well as phosphorylation-independent nucleocytoplasmic
shuttling. The group will also investigate how
these activities modulate the transcriptional functions of
the STATs and influence complex phenotypes such as
growth or antiviral protection.
The Microscopic Technology Group offers a range of conventional
and advanced light and electron microscopic
methods as well as electrophysiology and microinjection
to all interested research groups in the institute. The group
is available for all types of collaboration and the processing
of common research projects. So, e.g. in terms of characterizing
cyclic nucleotide-gated cation channels, it has
long standing collaborations with the Synthetic Organic
Biochemistry Group. This group has continued the development
of photolabile inactive compounds (caged com-
wie der Zugriff auf die jeweiligen Zielgene reguliert ist und
wie dies die Tanskriptionsprogramme beeinflusst. Das Ziel
der Gruppe ist die umfassende Beschreibung der Mechanismen,
nach denen der subzelluläre Transport der STATs
abläuft. Dabei legt sie besonderes Augenmerk auf die biochemischen
Grundlagen der Kernakkumulation und des
phosphorylierungsabhängigen nukleozytoplasmatischen
Transports. Zusätzlich wird untersucht, wie diese Aktivitäten
die Transkriptionsfunktionen der STATs modulieren und
komplexe Phänotypen, wie zum Beispiel Wachstum oder
antiviralen Schutz, beeinflussen.
Die Arbeitsgruppe Mikroskopische Techniken bietet allen
interessierten FMP-Gruppen ein breites Spektrum konventioneller
und spezialisierter licht- und elektronenmikroskopischer
Methoden sowie Elektrophysiologie und Mikroinjektion
an. Die Gruppe ist offen für Kooperationen und
gemeinsame Forschungsprojekte. Sie charakterisiert zum
Beispiel seit einigen Jahren zusammen mit der Arbeitsgruppe
Synthetische Organische Biochemie erfolgreich
durch zyklische Nukleotide getriebene Kationenkanäle.
Dabei kommen in dieser Arbeitsgruppe entwickelte, photolabile
chemische Verbindungen (caged compounds) zum
Einsatz, aus denen auf Anregung mit ultraviolettem Licht
hin aktive Biomoleküle freigesetzt werden. Diese Verbin-
pounds) from which the active biomolecules are liberated
by UV light. These compounds (caged cAMP, caged
cGMP) have proved extremely useful in elucidating intracellular
signal transduction chains.
The Cellular Physiology Group is concentrating on signal
transduction in the blood brain barrier (BBB), using co-cultures
of endothelial and glial cells as model systems. Current
emphasis is on the structure and function of tight
junction proteins sealing the interendothelial cleft. The
group is also investigating the role of protein kinases in the
regulation of the permeability of the BBB. The long-term
goal is to elucidate the molecular basis of such permeability
changes under physiological and pathological conditions.
The Biochemical Neurobiology Group is studying biochemical
and molecular interactions of peptide degrading
enzymes and their neuropeptide substrates under pathological
conditions. The research is e.g. focused on the
function of angiotensin-converting enzyme (ACE) with
alcohol reward in the CNS. This process involves angiotensin
receptors and their signal transduction pathways.
dungen (caged cAMP, caged cGMP) haben sich als außerordentlich
nützlich für die Untersuchung intrazellulärer
Signalketten erwiesen.
Die Arbeitsgruppe Zellphysiologie fokussiert ihre Forschung
auf die Signaltransduktion in der Blut-Hirn-Schranke
(BHS). Dabei verwendet sie gemeinsam kultivierte
Endothel- und Gliazellen als Modellsystem. Gegenwärtig
stehen insbesondere Struktur und Funktion von Tightjunction-Proteinen,
die den interendothelialen Spalt abdichten,
im Vordergrund des Interesses. Die Gruppe
erforscht auch die Rolle von Proteinkinasen bei der Regulation
der Durchlässigkeit der BHS. Ziel ist es, die molekulare
Basis der Veränderungen zu erforschen, denen die
Durchlässigkeit der BHS unter verschiedenen physiologischen
und pathologischen Bedingungen unterliegt.
Die Arbeitsgruppe Biochemische Neurobiologie untersucht
die biochemischen und molekularen Interaktionen
peptidabbauender Enzyme und ihrer Neuropeptidsubstrate
unter pathologischen Bedingungen. Die Forschung zielt
unter anderem auf die Rolle des Angiotensin-konvertierenden
Enzyms (ACE) und seiner Substrate beim Alkohol-
Reward im Belohnungssystem des ZNS. Dieser Prozess
verläuft unter Einbeziehung von Angiotensinrezeptoren
und deren nachgeschalteten Signalketten.

PROTEIN TRAFFICKING
Group Leader: PD Dr. Ralf Schülein
QUALITY CONTROL AND ER INSERTION OF
G-PROTEIN-COUPLED RECEPTORS
The group is interested in the early stages of the intracellular
trafficking of G-protein-coupled receptors (GPCRs).
The intracellular transport of membrane proteins starts
with their insertion into the membrane of the endoplasmic
reticulum (ER). During ER insertion, the proteins are folded,
and correct folding is monitored by a quality control
system (QCS). Only correctly folded proteins are allowed
to enter the vesicular transport via the ER/Golgi intermediate
compartment (ERGIC) and the Golgi apparatus to the
plasma membrane.
The study of the early stages of the intracellular transport
of GPCRs is of clinical significance. Naturally occuring
receptor mutations frequently lead to misfolded proteins
unable to pass the QCS and consequently to inherited
diseases. An example is X-linked nephrogenic diabetes
insipidus (NDI), which is caused by mutations in the gene
of the vasopressin V2 receptor (V 2R). It is noteworthy that
the QCS may work faultily. Sometimes, non-functional
mutant receptors are transported to the plasma membrane,
whereas in other cases, still functional proteins are
retained.
We use NDI-causing V 2R mutants to characterize the
mechanisms of GPCR quality control in the early secretory
pathway and to identify the proteins involved. Our aim is
to identify new drug targets for the treatment of diseases
caused by transport-deficient membrane proteins. In
addition, we study the function of cleavable signal peptides
of GPCRs.
1. Significance of cleavable signal peptides of
GPCRs
ER insertion of GPCRs is mediated by two different types of
signal sequences: one group contains signal anchor
sequences that are part of the mature receptors. A second
group possesses additional signal peptides that are cleaved
off during the insertion process by the signal peptidases
of the ER. The reason why this second subset requires
additional signal peptides was not clear for GPCRs and
other membrane proteins.
We studied the functional significance of the signal peptide
of the human endothelin B receptor and showed that
it does not influence receptor expression, but rather is
necessary for the translocation of the receptor’s N tail
across the ER membrane. We have now studied the signal
peptides of the rat corticotropin-releasing factor receptors
type 1 (CRF 1R) and type 2 α (CRF 2αR). The signal peptide
of the CRF 1R is not necessary for N tail translocation
but strongly promotes receptor expression. The signal
peptide of the CRF 2αR also promotes receptor expression.
However, it is not cleaved after ER insertion and remains at
the N tail of mature receptor. The CRF 2αR thus represents
the first GPCR containing eight hydrophobic helices: seven
transmembrane helices and an eighth helix with extracellular
location. The solubility of the latter helix seems to be
ensured by N-glycosylation.
2. Analysis of quality control mechanisms of
the V 2R
2.1. Quality control in the ERGIC
It was unknown whether quality control of membrane proteins
is restricted to the ER or whether the ERGIC or the
Golgi apparatus are also involved. We have addressed this
question using misfolded NDI-causing V2R mutants. Our
results show that these mutants fall into two classes. Class
A mutants (e.g. L62P, DL62-R64, S167L; Fig. 1) are retained
exclusively in the ER and never leave this compartment.
Class B mutants (e.g. R143P, Y205C, R337X, V226E, InsQ292;
Fig. 1), in contrast, reach the ERGIC. These mutants are
recognized in this compartment and are most likely rerouted
to the ER via the retrograde transport system. Moreover,
we could show that the ability of a mutant to reach
the ERGIC is not determined by its expression level but by
its folding state. The ERGIC may represent a second safety
net within the QCS recognizing those proteins that are only
inefficiently detected at the ER level.
2.2. Compartment-specific rescue of NDI-causing V 2R
mutants
Pharmacological strategies to rescue transport-deficient
mutant membrane proteins are of clinical significance.
Two strategies are being pursued at the moment. The first
involves the development of ligands that favor correct folding
of the mutant proteins and consequently allow them
to pass the QCS (“pharmacological chaperones”). The
second strategy is based on the fact that the QCS is sometimes
overprotective and retains mutant proteins that are
still functional. Here, substances that do not influence folding
but prevent the interaction with quality control components
may allow transport and lead to a functional rescue.
V 2R antagonists mediating the rescue of some NDI-causing
V 2R mutants as pharmacological chaperones have
been described. We have now found novel inhibitors of the
QCS acting in a compartment-specific way. By using the
L62P mutant (ER-retained; class A) and the Y205C mutant
(ERGIC-reaching; class B) of the V 2R as a model, we show
that the cell-penetrating peptide penetratin rescues the
Cellular Signalling/ Molecular Genetics
39

L62P
ΔL62-R64
transport of the Y205C mutant but not that of the L62P
mutant. We have also shown that the peptides exert their
function by displacing the BiP chaperone from the mutant
receptor in the ERGIC.
Group members
R143P
FIGURE 1
Two-dimensional model of the V2R. The positions of class A mutants (L62P, DL62-R64, S167L) are indicated by open rectangles, the positions of
class B mutants (R143P, Y205C, R337X, V226E, InsQ292) are indicated by black rectangles. The following posttranslational modifications of the V2R
are depicted: N-glycosylation at position N22, palmitoylation at positions C341 and C342 and a disulfide bond formed between residues C112 and
C192.
Dr. Ute Donalies
Dr. Claudia Rutz**
AnjaThielen (Doctoral student)*
Martina Alken (Doctoral student)*
Morad Oueslati (Doctoral student)
Dagmar Michl (Technical assistance)**
External funding
Deutsche Forschungsgemeinschaft
“Struktur und Funktion von Transportsignalen des Vasopressin
V2-Rezeptors” (Teilprojekt A3 im Sonderforschungsbereich
449 “Struktur und Funktion membranständiger
Rezeptoren”)
Ralf Schülein, Walter Rosenthal
* part of period reported
** part-time
S167L
Y205C
InsQ292
V226E
R337X
Deutsche Forschungsgemeinschaft
“ER-Insertion und Qualitätskontrolle von G-Protein-gekoppelten
Rezeptoren” (Teilprojekt A11 im Sonderforschungsbereich
366 “Zelluläre Signalerkennung und Umsetzung”)
Ralf Schülein, Walter Rosenthal
Selected publications (FMP authors in bold)
Oueslati M, Hermosilla R, Oorschot V, Donalies U,
Schönenberger E, Beyermann M, Oehlke J, Wiesner B,
Oksche A, Klumperman J, Rosenthal W, Schülein R (2005)
Compartment-specific rescue of nephrogenic diabetes
insipidus-causing vasopressin V2 receptor mutants by
cell-penetrating peptides. J Cell Biol, in revision
Thielen A, Oueslati M, Hermosilla R, Krause G, Oksche A,
Rosenthal W, Schülein R (2005) The hydrophobic amino
acid residues in the membrane-proximal C tail of the G protein-coupled
vasopressin V2 receptor are necessary for
transport-competent receptor folding. FEBS Lett, in press
Alken M, Rutz C, Köchl R, Donalies U, Oueslati M, Furkert
J, Wietfeld D, Hermosilla R, Scholz A, Beyermann M,
Rosenthal W, Schülein R (2005) The signal peptide of the
rat corticotropin-releasing factor receptor 1 promotes

L62P
Y205C
PE
PE
receptor expression but is not essential for establishing a
functional receptor. Biochem J 390, 455-64
Hermosilla R, Oueslati M, Donalies U, Schönenberger E,
Krause E, Oksche A, Rosenthal W, Schülein R (2004)
Disease-causing V2 vasopressin receptors are retained in
different compartments of the early secretory pathway.
Traffic 5, 993-1005
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G, Hermosilla
R, Schaefer M, Schülein R, Rosenthal W, Oksche
A (2004) Pharmacochaperones post-translationally enhance
cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2 receptors.
J Biol Chem 279, 47254-47263
Neuschäfer-Rube F, Rehwald M, Hermosilla R, Schülein R,
Rönnstrand L, Püschel G (2004) Requirement of different
Ser/Thr residues in the C-terminal domain of the human
EP4 receptor for agonist-induced phosphorylation‚ arrestin
interaction and sequestration. Biochem J 379, 573-585
Engelsberg A, Hermosilla R, Karsten U, Schülein R, Dörken
B, Rehm A (2003) The Golgi protein RCAS1 controls cell
surface expression of tumor-associated O-linked glycan
antigens. J Biol Chem 278, 22998-3007
2
M/I
0
2
M/I
0
PE
PE
Collaborations
FIGURE 2
Influence of penetratin treatment on the subcellular
distribution of the GFP-tagged V 2R mutants
L62P (class A) and Y205C (class B) in transiently
transfected HEK 293 cells. (A) Laser scanning
microscopy. Cells were treated with penetratin
(Pe, 1 µM) or with vehicle (-). The GFP fluorescence
signals of the receptors were recorded (green,
left panels), plasma membranes were stained with
Trypan blue (red, central panels) and overlay of the
signals was computed (right panels; co-localization
is indicated by yellow). Scale bars, 5 µm. (B)
Quantitative analysis of the peptide effect. The
ratio of cell membrane and intracellular fluorescence
signals (M/I) was determined. Ratios < 1
indicate a predominantly intracellular localization
of the receptors and ratios > 1 a predominant localization
at the cell membrane. Columns show mean
values ± SD (n = 30 cells). Note that in (A) and (B)
only the transport of the Y205C mutant is rescued
by penetratin treatment.
R.S. Hedge, NIH Bethesda, MD, USA
A. Oksche, Charité – University Medicine Berlin
R. Hermosilla, Charité – University Medicine Berlin
A. Rehm, Max Delbrück Center for Molecular Medicine
Berlin
G. Püschel, University of Potsdam
Cellular Signalling/ Molecular Genetics
41

ANCHORED SIGNALLING
Group Leader: PD Dr. Enno Klussmann
WATER REABSORPTION IN THE KIDNEY
A major function of the kidney is the production of urine. A
human kidney generates 180 l of primary urine per day,
most of which is water. It is obvious that excretion of such
vast amounts of fluid would cause dehydration and death.
Therefore, mechanisms have evolved which carefully
control body water balance by regulating water reabsorption
from primary urine. Most of the water (90%) is reabsorbed
by passive diffusion along an osmotic gradient
established between the primary urine and the surrounding
tissue. Reabsorption of the remaining 10% of water is
regulated by antidiuretic hormone (Arginine-vasopressin,
AVP) in particular cells of the kidney, the principal cells.
Principal cells line the collecting duct, the terminal part of
the tubular system transporting urine to the bladder. Loss
of responsiveness to the hormone results in a disease
known as diabetes insipidus (DI). DI is characterized by
a massive loss of water (up to 20 l per day if untreated).
The molecular mechanism underlying AVP-mediated
water reabsorption in the kidney involves several key proteins
including the vasopressin V2 receptor, and aquaporin-2
(AQP2). The latter belongs to the family of water
channels whose discovery yielded Peter Agre the Nobel
Prize in Chemistry in 2003. AVP binds to the vasopressin
V2 receptor located on the surface of renal principal cells
thereby triggering the redistribution of AQP2 from intracellular
vesicles into the apical (urine-facing) plasma
membrane. This process is also known as the AQP2
shuttle.
The insertion of AQP2 into the plasma membrane introduces
water-selective pores and facilitates water entry into
the cells. Water exits the cells through aquaporins-3 and
-4 constitutively located in the basolateral (tissue-facing)
plasma membrane. The driving force is again the osmotic
gradient established between primary urine and the tissue.
On the molecular level, AVP stimulates the elevation
of the second messenger cAMP followed by activation of
protein kinase A (PKA). PKA, in turn, transfers a phosphate
group to AQP2. This phosphorylation is a prerequisite
for the redistribution of AQP2. The mechanism is depicted
in Figure 1.
The aim of our group is to identify further proteins involved
in the redistribution of AQP2. This will lead to a detailed
understanding of the underlying mechanism, and may
eventually lead to a treatment of certain cases of DI. The
translocation of AQP2 from intracellular vesicles to the
plasma membrane constitutes an exocytosis-like process.
A better understanding of this mechanism will also yield a
better understanding of other cAMP-dependent exocytic
processes such as renin secretion from juxtaglomerular
cells, insulin secretion from pancreatic βcells or proton
secretion from gastric parietal cells. Dysregulations of
these processes cause various diseases including hypertension,
diabetes mellitus and gastric ulcers.
AKAP18δ anchors PKA to AQP2-bearing vesicles
The above mentioned phosphorylation of AQP2 is not the
sole prerequisite for its redistribution. We have shown that
the tethering of PKA to subcellular compartments by so
called A-kinase anchoring proteins (AKAPs) is also essential
for the AQP2 shuttle to occur. During the search for
AKAP(s) involved in the shuttle, a new splice variant of
AKAP18, AKAP18δ, was identified. Biochemical and cell
biological approaches revealed that AKAP18δ functions
as an AKAP in vitro and in vivo. Immunofluorescence
microscopy showed that in the kidney, AKAP18δ is mainly
expressed in principal cells of the terminal section of the
collecting duct, closely resembling the distribution of
AQP2. AKAP18δ was identified on the same intracellular
vesicles as AQP2 and PKA. AVP not only recruited AQP2,
but also AKAP18δ to the plasma membrane (Fig. 2). AVP
caused the dissociation of AKAP18δ and PKA. The data
suggest that AKAP18δ is involved in the AQP2 shuttle by
anchoring PKA in close proximity to AQP2 (Henn et al.
2004).
The small GTPase RhoA mediates the diuretic
effects of prostaglandin E2
The small GTPases of the Rho family (Rho, Rac, Cdc42)
participate in the regulation of the F-actin cytoskeleton. In
previously published papers we have demonstrated that
the small GTPase RhoA, a particular member of the Rho
family, in its active state (GTP-bound) induces the formation
of F-actin-containing stress fibers in primary cultured
principal cells,and prevents the AQP2 shuttle. We observed
that AVP causes a decrease of F-actin-containing
stress fibers, and inhibition of RhoA.
Prostaglandin E 2 (PGE 2) antagonizes AVP-induced water
reabsorption. Using primary cultured rat inner medullary
collecting duct principal (IMCD) cells, we have shown that
stimulation of prostaglandin EP 3 receptors induced RhoA
activation and formation of F-actin-containing stress fibers
in resting IMCD cells, but did not modify the intracellular
localization of AQP2. However, the AVP-induced AQP2
translocation was strongly inhibited. In addition, stimulation
of EP 3 receptors inhibited the AVP-induced RhoA in-

A B
FIGURE 1
Schematic representation of the vasopressin-mediated water reabsorption. In resting principal cells (A) aquaporin-2 (AQP2) is located in intracellular
vesicles. B. The binding of arginine-vasopressin (AVP) to the vasopressin V2 receptor (V 2R) activates a cAMP-dependent signalling cascade
which causes the phosphorylation of AQP2 (phosphorylated AQP2, p-AQP2) and its translocation from intracellular vesicles into the plasma
membrane facing the lumen of the renal collecting duct. Water from the primary urine enters the cells through AQP2 along an osmotic gradient
and exits the cells through aquaporin-3 and aquaporin-4 (AQP3 und AQP4) constitutively present in the basolateral plasma membrane. AC, adenylyl
cyclase; G s, stimulatory G-Protein; C and R, catalytic and regulatory subunits of protein kinase A (PKA), respectively.
activation and the AVP-induced depolymerization of
F-actin-containing stress fibers. The inhibitory effect of EP 3
receptor stimulation was independent of increases in
cAMP and cytosolic Ca 2+ and is most likely mediated by
the G-proteins G12/13. Further experiments showed that
elevation of cAMP results in the phosphorylation of RhoA
by PKA, which is known to inhibit this GTPase. Taken
together, the data suggest that the signaling pathway
underlying the diuretic effects of PGE 2 (and probably those
of other diuretic agents) include cAMP- and Ca 2+-independent
RhoA activation and F-actin formation (Tamma et al.
2003a, 2003b).
Group members
Dr. Dorothea Lorenz**
Dr. Theresa McSorley
Dr. Volker Henn
Dr. Pavel Nedvetsky
Katja Santamaria (Doctoral student)
Viola Weber (Doctoral student)
Eduard Stefan (Doctoral student)
Christopher Blum (Doctoral student)
Christian Hundsrucker (Doctoral student)
** part-time
Andrea Geelhaar (Technical assistance)
Michael Gomoll (Trainee)
External funding
Deutsche Forschungsgemeinschaft
„Die Rolle des Zytoskeletts bei der Vasopressin-induzierten
Translokation von Aquaporin-2 in die Plasmamembran
renaler Hauptzellen“ (Kl1415/1-1)
E. Klussmann, W. Rosenthal
Deutsche Forschungsgemeinschaft
„Charakterisierung der Proteinkinase A-Ankerproteine
Ht31 und Rt31 und Untersuchungen zu ihrer biologischen
Funktion“ (Ro597/9-1)
W. Rosenthal, E. Klussmann
European Community, 5th Frame work programme
“Anchored cAMP signalling - implications for treatment of
human disease” (QLK3-CT-2002-02149)
E. Klussmann, W. Rosenthal
Cellular Signalling/ Molecular Genetics
43

-AVP
+AVP
AQP2 AKAP18δ overlay
Selected publications (FMP authors in bold)
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W,
Pohl P, Maric K (2003) Cyclic AMP is sufficient for triggering
the exocytic recruitment of aquaporin-2 in renal epithelial
cells. EMBO Rep 4, 88-93
Tsunoda SP, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin-1, nothing but a water channel. J Biol
Chem 279, 11364-11367
Tamma G, Wiesner B, Furkert J, Oksche A, Schaefer M,
Valenti G, Rosenthal W, Klussmann E (2003) The prostaglandin
E2 analogue sulprostone antagonizes vasopressin-induced
antidiuresis through activation of Rho. J Cell
Sci 116, 3285-3294
Tamma G, Klussmann E, Procino G, Svelto M, Rosenthal
W, Valenti G (2003) cAMP-induced AQP2 translocation is
associated with RhoA inhibition through RhoA phosphorylation
and interaction with RhoGDI. J Cell Sci 116, 1519-
1525
Storm R, Klussmann E, Geelhaar A, Rosenthal W, Maric K
(2003) Osmolality and solute composition are strong regulators
of AQP2 expression in renal principal cells. Am J
Physiol 284 (Renal Section), F189-198
Collaborations
FIGURE 2
The effect of AVP on the subcellular distribution of
AQP2 and AKAP18δ in primary cultured rat inner
medullary collecting duct principal (IMCD) cells. IMCD
cells were left untreated (-AVP) or incubated with AVP
(100 nM, 15 min), fixed and permeabilized. AQP2 was
detected by incubation with goat anti-AQP2 and
secondary Cy3-conjugated antibodies, AKAP18δ with
affinity-purified rabbit anti-AKAP18δ A18δ4 and
secondary Cy5-conjugated antibodies. Immunofluorescence
signals were detected by laser scanning
microscopy. The overlay of Cy3 and Cy5 fluorescence
signals is shown in the right panel. Scale bars, 20 µm.
Prof. K. Tasken, University of Oslo, Norway
AKAPs in cardiac myocytes
Dr. M. Zaccolo, University of Padova, Italy
Modulation of Ca2 + fluxes in cardiac myocytes
Professor J. D. Scott, Howard Hughes Medical Institute,
Vollum Institute, Oregon Health & Sciences University,
Portland, Oregon, USA
Peptide disruptors of AKAP-PKA interactions

Prof. M. Houslay, University of Glasgow, Scotland, UK
Role of phosphodiesterases in renal principal cells
Prof. G. Valenti, University of Bari, Italy
Rho-dependent signalling in the AVP-mediated water
reabsorption
Professor F. W. Herberg, University of Kassel
Analyses of AKAP-PKA interactions
Cellular Signalling/ Molecular Genetics
45

CELLULAR IMAGING
Group Leader: Dr. Burkhard Wiesner
ENHANCEMENT OF THE CELL SURFACE
EXPRESSION BY INCREASING CONFOR-
MATIONAL STABILITY OF VASOPRESSIN
V2 RECEPTORS
This facility offers a range of conventional and advanced
light and electron microscopic methods as well as electrophysiology
to all interested research groups in the institute.
The group is available for all types of collaboration
including advice in preparation techniques, and the processing
of common research projects.
About 50% of nephrogenic diabetes insipidus (NDI)-causing
mutations in the vasopressin V 2 receptor (V 2R) gene
code for single amino acid replacements (missense mutations).
In most cases the encoded mutant V 2Rs are misfolded
and retained in the ER. Improvement of this situation
is to be achieved by the application of specific substances
(e.g. pharmacochaperones). Such cell permeable antagonists
should restore cell surface expression. So we
studied the effects of SR121463B and SR49059 (V 2R and
V 1R-specific antagonists) on ER-retained V 2Rs transiently
expressed in HEK293 cells. Such investigations can be
accomplished generally by receptor binding studies
(radioactive proof of the number of receptors of the cell
surface). If, however, the assigned substance (pharmacochaperone)
blocks the binding domain for the ligand, this
method is not applicable. In this situation, the employment
of confocal microscopy is very helpful. So the receptor
can be located with the help of green fluorescent protein
(GFP) microscopically. Optical staining of the cell membrane
by further coloring (trypan blue) makes a simple allocation
possible between intracellular and membrane localization
of the receptor (see Fig. 1).
In the case of the wild-type murine V 2R (mV 2R), which is
mainly localized in the ER in untreated controls, both antagonists
promoted a time- and dose- dependent restoration
of the cell surface expression. SR49059-mediated
restoration of cell surface expression of the mV 2R was
accompanied by a dramatic increase in [ 3H]AVP binding
sites.
Figure 2 shows, that with the microscopic investigation the
rearrangement (from the ER to the cell membrane) is fast
run off. This phenomenon is justified by the optical resolution
of light microscopy. The binding studies show the
functional receptor in the plasma membrane of the cell.
Light-microscopy cannot differentiate whether the receptor
is located at the intracellular side of the membrane or
in the plasma membrane of the cell.
For human NDI-causing mutants (hL62P, hΔLAR 62-64,
hH80R, hW164R, hS167T, hS167L, hC319Y, hP322S) and
several in vitro mutants (hD136A, hD368K/S371X, hF328A)
transiently expressed in HEK cells, also predominant ER
retention was observed. Interestingly, cell surface expression
of hΔLAR 62-64, hD136A, hS167T, hP322S, and hF328A
was only restored by SR121463B. In the case of the
mutants hL62P, hH80R, hW164R, hS167L, and hD368K/
S371X none of the two antagonists restored cell surface
expression. Only for the mutant hC319Y, both antagonists
promoted cell surface expression as found for the mV 2R.
The data show that ER-retained mutant V 2Rs differ in their
sensitivity to antagonist-promoted cell surface expression.
It is likely that these differences can be attributed to the
extent of the folding defect and/or an alteration of the binding
pocket. Further studies are required to analyze the
general applicability of antagonists in the treatment of NDI.
Further projects involved:
A-kinase anchoring proteins and the exocytotic recruitment
of aquaporin-2 (D. Lorenz, B. Wiesner, M. Ringling, B.
Oczko in cooperation with Enno Klussmann, Anchored
Signalling)
Application of novel caged compounds (B. Wiesner, J.
Eichhorst, B. Oczko in cooperation with Volker Hagen, Synthetic
Organic Biochemistry)
Water flux through water channels (D. Lorenz, B. Wiesner,
B. Oczko in cooperation with Peter Pohl, Biophysics)
Cellular uptake of peptides (B. Wiesner, B. Oczko in cooperation
with J. Oehlke, Peptide Lipid Interaction/Peptide
Transport; and M. Beyermann, Peptide Synthesis)
Constitutive internalization of the human V2 Vasopressin
receptor (A. Schmidt, B. Wiesner in cooperation with R.
Hermosilla, Charité group)
DNA binding controls inactivation and nuclear accumulation
of the transcription factor Stat1 (B. Wiesner, B. Oczko
in cooperation with U. Vinkemeier, Cellular Signal Processing)
Enhancement of the cell surface expression by increasing
conformational stability of G protein coupled receptors
(B. Wiesner, J. Eichhorst, B. Oczko in cooperation with
A. Oksche, Charité group; and S. Wüller, RWTH Aachen
Medical School).

Trypan blue mV2R.GFP
A C C
FIGURE 1
HEK293 cells transiently expressing the mV 2R.GFP were treated with the vasopressin receptor antagonist SR49059 for up to 13 h (A: oh; B: 7 h;
c: 13 h). Top panel: mV 2R.GFP fusion protein. Bottom panel: Plasma membrane stained with trypan blue.
FIGURE 2
Antagonist-mediated restoration of cell surface expression analyzed by quantitative laser scanning microscopy and binding analysis. Cells transiently
expressing the mV 2R.GFP were treated for up to 16 h with the specific vasopressin receptor antagonist. The fitted curve representing the
increase in the normalized fluorescence intensities is shown in black. In parallel, membrane preparations were analyzed for specific binding of
[ 3H]AVP. The curve representing the increase in specifically bound [ 3H]AVP is shown in gray.
Cellular Signalling/ Molecular Genetics
47

Group members
Dr. Dorothea Lorenz**
Antje Schmidt (Student)*
Brunhilde Oczko (Technical assistance)
Jenny Eichhorst (Technical assistance)*
Martina Ringling (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Degradations-Mechanismen des humanen Vasopressin-
V2-Rezeptors und einiger von Patienten mit X chromosomaler
nephrogener Diabetis insipidus isolierten V2-
Rezeptormutanten“ (He 4486/1-1 und 1-2)
Ricardo Hermosilla (Charité - University Medicine Berlin),
Burkhard Wiesner
Selected publications (FMP authors in bold)
Geissler D, Kresse W, Wiesner B, Bendig J, Kettenmann
H, Hagen V (2003) DMACM-Caged Adenosine Nucleotides:
Ultrafast Phototriggers for ATP, ADP, and AMP Activated
by Long-Wavelength Irradiation. ChemBioChem 4, 162-170
Hagen V, Frings S, Wiesner B, Helm S, Kaupp UB, Bendig
J (2003) [7 (Dialkylamino)-coumarin 4 yl]methyl-caged
compounds as ultrafast and effective long-wavelength
phototriggers of 8 Bromo-substituted cyclic nucleotides.
ChemBioChem 4, 434-442
Meyer T, Marg A, Lemke P, Wiesner B, Vinkemeier U
(2003) DNA binding controls inactivation and nuclear
accumulation of the transcription factor Stat1. Genes Dev
17, 1992-2005
Tamma G, Wiesner B, Furkert J, Hahm D, Oksche A,
Schaefer M, Valenti G, Rosenthal W, Klussmann E (2003)
The prostaglandin E2 analogue sulprostone antagonizes
vasopressin-induced antidiuresis through activation of
Rho. J Cell Sci 116, 3285-3294
Tsunoda S, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin 1, nothing but a water channel. J Biol
Chem 279, 11364-11367
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin 2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
* part of period reported
** part-time
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G, Hermosilla
R, Schaefer M, Schülein R, Rosenthal W, Oksche
A (2004) Pharmacochaperones post-translationally enhance
cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2 receptors.
J Biol Chem 279, 47254-47263
Collaborations
Prof. Ricardo Hermosilla
Pathology of signal transduction
Charité-Universitätsmedizin Berlin
PD Dr. Alexander Oksche
G protein-coupled receptors: Signal transduction and
regulation of cell surface expression
Charité-Universitätsmedizin Berlin

MOLECULAR CELL PHYSIOLOGY
Group Leader: Dr. Ingolf Blasig
STRUCTURE, FUNCTION, AND REGULATION
OF CELL-CELL CONTACT PROTEINS
This group investigates signal transduction pathways
including cell-cell and protein-protein interactions under
normal and pathological conditions, such as oxidative
stress in the brain. The aim of these studies is to explore
molecular aspects of structure, function and regulation of
blood-brain barrier (BBB) proteins for the development of
new pharmacological approaches to improve treatment
of cerebral diseases, such as stroke, inflammation,
epilepsy etc. Another aspect is to develop approaches to
open the BBB for pharmacokinetic application, e. g. improved
administration of such antitumor or antiepileptic drugs
which are non-permeable through the BBB.
A main focus was the self-association mechanism of
transmembrane and membrane associated proteins forming
the tight junctions (TJ). The regulatory and transmembrane
TJ protein occludin was found colocalizing within
cell membrane contacts of the same cell and could be
coprecipitated with itself (intracellular association).
Differently tagged TJ strand-forming claudin-5 also colocalized
in the cell membrane of the same cell and showed
fluorescence resonance energy transfer (intracellular
association). This demonstrates self-association both of
occludin and of claudin-5 in one cell membrane. For occludin,
dimerization of the cytosolic C-terminal coiled coildomain
was identified. In claudin-5, we detected that the
second extracellular loop is a dimer. Thus, occludin may
self-associate via its coiled coil-domain and claudin-5 via
its second extracellular loop. Since the transmembrane
junctional adhesion molecule JAM is known to dimerize in
TJ, we hypothesize that homodimerization is a structural
feature of transmembrane TJ proteins. This potential is
corroborated by our finding that the recruiting protein of
the transmembrane TJ proteins, the membrane-associated
ZO-1 also forms a dimer. A general dimerization principle
of transmembrane TJ proteins via the ZO-1 has been
worked out. This principle may serve as a common feature
of TJ assembly (Figure). This work was supported by
groups of the FMP (G. Krause, Structural Bioinformatics;
E. Krause, Mass Spectrometry) and MDC (K. Gast; M.
Kolbe).
The occludin-ZO-1 interaction was studied in more detail.
Binding studies by SPR and peptide mapping combined
with CD studies and molecular modelling indicated that
occludin’s coiled-coil domain interacts as a three-helix
bundle with three helices on the SH3-hinge-GuK unit of
ZO-1. A similar association was found between ZO-1 and
the adherens junction protein α-catenin. Ion bindings between
the basic helices of ZO-1 and the acidic ones in
occludin were also identified. In conclusion, a common
molecular mechanism of forming intermolecular helical
bundles between ZO-1 and α-catenin and occludin was
identified as a general molecular principle organizing the
association of ZO-1 at adherens and tight junctions. Collaboration
within the FMP (G. Krause, Structural Bioinformatics;
M. Beyermann, Peptide Chemistry), and with the
Free University (O. Huber) and the Humboldt University
(J. Schneider-Mergener) Berlin.
Further studies were accomplished on the effects of hypoxia
on cultures of brain capillary endothelial cells (BCEC).
In addition to identifying proteins that are up- or downregulated
after hypoxia, activity data were acquired for
selected proteins. Novel proteomic techniques have been
introduced which allow the elucidation of signalling pathways
by analyzing post-translational modifications (e. g.,
protein phosphorylation). Moreover, investigations were
aimed at improving the conditions for targeting low abundant
proteins using the Rotofor © technology (collaboration
E. Krause, Dept. Peptide Chemistry; D. Stanimirovic/Ottawa,
Canada) as well as for membrane proteins. Earlier
results suggested that cytokines are involved in the
response to oxidative stress. Continuative investigations
showed that exposure to interleukin-1ß results in proliferation
of the cells, nerve growth factor (NGF) release and
expression of NGF receptors. This is a new pathway in
BCEC which, hence, can modulate BBB functions (collaboration
C. Humpel, Innsbruck/Austria).
Clinical studies were continued to support our experimental
findings that oxidative stress may injure the BBB and
that these disturbances can be treated. Thus, mood disorders
with depression were studied and indications were
found for opening of the BBB. BBB disturbances and schizophrenic
events were reduced by pharmacological
approaches targeted, among others, to astrocytes maintaining
BBB properties in the barrier forming brain endothelium
(collaboration M. Schroeter, Max Planck Institute,
Leipzig).
Group members
Dr. Reiner F. Haseloff
Dr. Jörg Piontek
Dr. Christine Rückert*
Dr. Darkhan Utepbergenov*
Anna Y. Andreeva (Doctoral student)*
* part of period reported
Cellular Signalling/ Molecular Genetics
49

Claudin-5 dimer JAM dimer Occludin dimer
FIGURE 1
Scheme of the dimerization concept showing that transmembrane TJ proteins, as well as ZO-1 may dimerize. Claudin-5 dimerizes via its second
extracellular loop (2.ECL; solid double arrow) and occludin via its coiled coil-domain (CC; interacting cylinders). JAMs are known to dimerize via
its extracellular domain (asterisk). ZO-1 dimerizes via its SH3-GuK unit. The positively charged (+) helices of ZO-1 (H1, H2 in GuK; CC1 between
SH3 and GuK) bind to negatively charged (-) CC-domain of occludin (two dotted arrows). PDZ-domain 3 of ZO-1 can associate to JAMs, PDZ-1
to claudins. N, N-terminus; yellow circles, disulfide bridge in claudin-5 first and occludin last loop.
Manjot Singh Bal (Doctoral student)
Dörte Lorberg (Doctoral student)*
Kerstin Mikoteit (Doctoral student)*
Sebastian L. Müller (Doctoral student)
Juliane Walter (Doctoral student)*
Lars Winkler (Doctoral student)
Inga Roswadowski* (Student)
Jenny Kirsch (Student)*
Markus Heine (Student)*
Birgit Lassowski (Student)*
Christian Niehage (Trainee)*
Katrin Schulz (Student)*
Ariane Schuster (Student)*
Constanze Wolf (Student)*
Nikolaj Zuleger (Student)*
Barbara Eilemann (Technical assistance)
Gislinde Hartmann (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Extrazelluläre Loops von Blut-Hirnschranken-Proteinen“
(BL 308/7-1)
Ingolf Blasig
* part of period reported
PDZ domains
SH3 domain
ZO-1 dimer
cell membrane
GuK domain
extra cellular
Deutsche Forschungsgemeinschaft
„Wechselwirkungen von Blut-Hirnschranken-Proteinen“
(BL 308/6-1, 6-2, 6-3)
Ingolf Blasig, Gerd Krause
Deutsche Forschungsgemeinschaft
„Lokalisation und Phophorylierungsmuster von Occludin“
(GK 238/3, Teilprojekt im Graduiertenkolleg „Schadenmechanismus
im Nervensystem – Einsatz von bildgebenden
Verfahren“
Ingolf Blasig
Bundesministerium für Bildung und Forschung
„Molecular Fingerprinting of the Blood-Brain Barrier in
Hypoxia – Targeting Brain Vessels to Treat Stroke“, NRC
Reiner Haseloff
Deutscher Akademischer Austauschdienst
„Molecular pharmacology of the HISS-resistin system”
(PPP-Ungarn 324/ssch)
Ingolf Blasig
Deutscher Akademischer Austauschdienst
„Natural polyphenols in the cardiovascular system”
(323/bis Slowakei)
Ingolf Blasig

European Community
Marie Curie Grant (HPMT-CT-2001-00399)
G. Schreibelt
Schering-Stiftung
„Phosphorylierung von Occludin” (Stipendium)
Anna Andreeva
Selected publications (FMP authors in bold)
Müller SL, Portwich M, Schmidt A, Utepbergenov DI,
Huber O, Blasig IE, Krause G (2005) The tight junction protein
occludin and the adherens junction protein β-catenin
share a common interaction mechanism with ZO-1. J Biol
Chem, in press
Zassler B, Blasig IE, Humpel C (2005) Protein delivery of
caspase-3 induces cell death in malignant C6 glioma and
brain capillary endothelial cells. J Neuro-Oncology, in
press
Haseloff RF, Blasig IE, Bauer H-C, Bauer H (2005). In
search of the astrocytic factor(s) modulating blood-brain
barrier functions in brain capillary endothelial cells in vitro.
Mol Cell Neurobiol 25, 25-39
Schmidt A, Utepbergenov DI, Mueller SL, Beyermann M,
Schneider-Mergener J, Krause G, Blasig IE (2004) Occludin
binds to the SH3-hinge-GuK unit of zonula occludens
protein 1 - potential mechanism of tight junction regulation.
Cell Mol Life Sci 61, 1354-1365
Moser KV, Reindl M, Blasig IE, Humpel C (2004) Brain
capillary endothelial cells proliferate in response to NGF,
express NGF receptors and secrete NGF after inflammation.
Brain Res 1017, 53-60
Schroeter ML, Abdul-Khaliq H, Frühauf S, Höhne R, Schick
G, Diefenbacher A, Blasig IE (2003) Serum S100B is increased
during early treatment with antipsychotics and in deficit
schizophrenia. Schizophrenia Res 62, 231-236
Collaborations
Hartwig Wolburg, Universität Tübingen
Wolf-Hagen Schunck, Max-Delbrück-Center für Molekulare
Medizin Berlin-Buch, Germany
Marina Bigl, Universität Leipzig, Germany
Hannelore Haase, Max-Delbrück-Center für Molekulare
Medizin Berlin-Buch, Germany
Danica Stanimirovic, Institute of Biological Sciences, NRC,
Ottawa, Canada
Hans-Christian Bauer, Universität Salzburg, Austria
Maria Balda, University College London, UK
Christian Humpel, Universität Innsbruck, Austria
Stephan Christen, Universität Bern, Switzerland
Cellular Signalling/ Molecular Genetics
51

BIOCHEMICAL NEUROBIOLOGY
Group Leader: Dr. Wolf-Eberhard Siems
UNEXPECTED FUNCTIONS OF PEPTIDOLY-
TIC ENZYMES
The Biochemical Neurobiology Group deals with the biochemical,
pharmaceutical and molecular aspects of peptidases.
The current research is focused on angiotensinconverting
enzyme (ACE), neutral endopeptidases (NEP)
and some related enzymes. We evaluate their biochemical
and functional relations to various disorders in
humans, like alcohol addiction, obesity, neuronal disorders,
problems in male fertility and heart diseases.
Peptidases play a frequently underestimated role in maintaining
and controlling essential functions in the body.
Anthony Turner, one of the Nestors of research on NEP,
wrote: “Peptidases and peptidolysis play vital roles in cellular
processes from fertilization – to death”. A look at the
ACE confirms this sentence: This enzyme plays a key role
not only for regulation of blood pressure, but also at the
beginning and at the end of life: it is essential for mammalian
fertilization (male ACE-knock out mice are infertile in
spite of normal sperm and libido) as well as for apoptosis,
in which angiotensin II (Ang II, the main product of ACE) is
essentially involved.
In recent years, the use of molecular techniques and of
genetically modified animals in peptidase research resulted
in tremendous progress and innovative perspectives.
In 2003 A. Turner commented the newly found functions of
ACE and NEP as follows: “... (they) provide new avenues
for the treatment of some of the major human diseases of
the aging population of the Western world: cardiovascular
disease, cancer and dementia”.
ACE, NEP and related enzymes are transmembranal
metallopeptidases, which are prevalent in many tissues
and cleave a broad spectrum of endogenous substrates.
They consist of a short cytoplasmatic domain, a single
transmembranal part and a very big extracellular part,
which includes the catalytic domain(s), each of them containing
one zinc ion.
ACE and NEP play an important role in circulation, and
inhibitors of the two enzymes, especially ACE inhibitors,
are widely used for treatment of hypertension and for
cardioprotection. Because of their broad spectrum of
other substrates, these enzymes are also significantly
involved in many other pathophysiological processes.
In recent years, a lot of novel, in part surprising results
were published on ACE and NEP. One example concerns
ACE2, an enzyme closely related to ACE: in addition, to its
peptidolytic activity, ACE2 was identified as the essential
receptor of the SARS virus, and represents the entry portal
of the body for the virus.
Kohlstedt et al. (2002) described receptor-like functions
also for ACE. They proved e.g. that the reaction of ACE with
some, but not all substrates or inhibitors resulted in specific
phosphorylation at its own cytoplasmatic site.
Another unexpected finding: the two enzymes, especially
the NEP, play an important role in degrading the Alzheimer’s
disease (AD) peptide ß-amyloid (Aß). This knowledge
is expected to lead to new basic approaches for
understanding and treatment of this disease.
In the Biochemical Neurobiology Group additional functions
of these peptidases have been discovered during the
last years.
ACE and voluntary alcohol consumption
Our experiments of the last years had demonstrated that
Ang II, the main product of ACE, directly influences voluntary
consumption of alcohol. Open questions concern the
role of central and peripheral Ang II, the involved receptor(s)
and the downstream signaling.
We characterized the role of central Ang II in alcohol intake
by using transgenic rats [TGR(ASrAOGEN)680], which
express an antisense RNA against angiotensinogen and
consequently have sharply reduced Ang II levels exclusively
in the central nervous system. These rats consumed
markedly less alcohol in comparison to their wild-type
controls. Moreover, Spirapril, an ACE inhibitor, which passes
the blood-brain barrier, did not alter voluntary alcohol
consumption in the TGR(ASrAOGEN)680, but it significantly
reduced alcohol intake in wild-type rats. Studies in different
types of knockout mice have proved that the effect of
Ang II on alcohol consumption is mediated by the angiotensin
receptor AT1, whereas the AT2 receptor and the
bradykinin B2 receptor are not involved. With reference to
signal transduction we found that the TGR(ASrAOGEN)680
with the very low central Ang II showed a markedly reduced
dopamine concentration in their ventral tegmental
area (VTA), confirming a role of dopaminergic transmission
in Ang II-controlled alcohol preference. Our results indicate
that a distinct drug-mediated control of the central
renin-angiotensin system (RAS) could be a new principle
for therapy of alcohol disease.
NEP-deficient mice – a model of human obesity
In long-lasting experiments on the functions of NEP, we
observed that elderly NEP-deficient mice (NEP-/-) had a
considerably higher body weight than wild-types. In con-

Recovery of ANP (%)
100
80
60
40
20
Mouse ANP
10 20 30 40 50 60
Incubation time (min)
NEP
FIGURE 1
Catabolism of murine ANP and BNP by lung membranes of wild-type and NEP-knockout mice: NEP is involved in the cleavage of ANP only.
A large percentage of the ANP-degradation and the total BNP-degrading activity (green arrows) cannot be ascribed to NEP (black arrow). An
interesting result is the upregulation of the non-NEP activity (yellow arrows) in NEP-deficient membranes, so that the degradation is faster than
in wild-type membranes.
trast to other animal models of obesity, but in accordance
with typical human obesity, the differences in body weight
became most apparent in the second half of life. NMR-studies
showed that the higher body weight in NEP-/- mice is
exclusively due to an accumulation of fat. The molecular
basis of NEP-related obesity in mice and the pathophysiological
consequences are presently being investigated.
NEP is known to hydrolyze a great number of hormones,
among them orexigenes (peptides stimulating food intake),
like NPY, MCH and some of the opioids. Furthermore, several
other peptides with effects on food ingestion also contain
amino acid sequences that should be hydrolyzed by
NEP (e.g. galanin, ghrelin, orexin). The catabolic effect of
NEP on these peptides and a more detailed biochemical
analysis of old NEP-/- mice with regard to a new obesity
model are still being investigated at present.
In contrast to other genetically modified animals expressing
obese phenotypes, the alterations in NEP-/- mice concern
a relatively great number of substrates and receptor
systems. Therefore, these mice may be a better animal
model of the typical, polyfactorial human obesity. These
results and derived diagnostic and therapeutic applications
are summarized in a joint patent application of FMP,
Charité und Free University of Berlin.
10 20 30 40 50 60
Incubation time (min)
� wild-type � wild-type + Candoxatrilat � NEP -/- � NEP -/- + Candoxatrilat � heat-inactivated wild-type
Recovery of BNP (%)
100
80
60
40
20
Mouse BNP
NEP deficiency in mice: learning experiments
NEP belongs to the enzymes which are able to catabolize
the amyloid ß-peptide (Aß). This important function of NEP
was recently confirmed by several in vitro as well as in
vivo studies and is now being intensively discussed as to
its impact in the pathogenesis and therapy of AD.
But in even very old NEP-/- animals immunostaining experiments
with brain slices and monoclonal antibodies
against murine Aß-peptide (cooperation with Multhaup,
Heidelberg/Berlin) did not reveal any Aß depositions.
Therefore, further experiments focused on differences in
the NEP-induced degradation of human and murine Aß or
Aß-partial sequences by using purified enzyme as well as
membrane preparations of NEP-/- and NEP+/+ mice. Aß
peptides of humans and rodents had indeed different
degradation rates (mAß>hAß). However, the differences
in Aß degradation by NEP cannot be the only reason for
the general lack of AD-like processes in rodent brains. Our
degradation studies indicated that further membrane
enzymes are involved in Aß catabolism. Surprisingly, also
ACE degraded Aß by means of an endopeptidolytic attack.
However, we found in behavioral experiments that the very
old homozygous NEP-knockout mice (>2 years) displayed
Cellular Signalling/ Molecular Genetics
53

A
B
cavity
catalytic site
an even better performance in learning tests, e.g. in a Morris
water maze, compared to their wild-type littermates.
This result was confirmed in long-term potentiation (LTP)
experiments in two brain regions (hippocampus and amygdala).
In contrast, there were no differences between the
two genotypes in younger animals (~9 months). The molecular
background of this effect is still unclear. These
experiments demonstrate the Janus head-like action of
NEP in brain function: it improves the brain function by
degradation of the plaque-forming Aß-peptides, and it
apparently degrades other peptides which improve learning
processes. Involved peptides are still unknown. Glucagon-like
Peptide-1 (GLP-1) could play a role. This peptide
strengthens cognitive functions, and we found a greater
stability of this peptide in membrane preparations of
NEP-/-- mice.
Domain-selective forms of ACE
To analyze the newly discovered ACE-functions and the
unexpected interactions with substrates and inhibitors in
more detail, we transfected CHO cells with the following
murine constructs: a) wild-type ACE (both domains are
intact), b) C-terminal ACE, c) N-terminal ACE, d) inactivated
ACE (no intact domain). The specificity of these ACE-
two large tails:
spatial clashes
FIGURE 2
A: Three-dimensional structure of the catalytic centre of
NEP.
B: Hypothesis of the NEP – NP interaction.
• NP moves into the cavity of NEP
• Complementary recognition sites are supporting the orientation
of NP within the cavity
• Large N - and C - terminal tails (simultaneously) hinder
the orientation towards the catalytic site
forms was characterized by Western blots and enzymatic
studies with domain selective substrates (Hip-His-Leu,
Z-Phe-His-Leu, Ac-SDKP), and the clones each with the
highest enzyme activities were selected.
These different domain-selective ACE forms are now used
to investigate
(i) the receptor-like function of cell-bound ACE,
(ii) the unusual endopeptidolytic activity of ACE on Aß and
Aß partial sequences,
(iii) the role of angiotensin (1-7) as substrate and inhibitor.
Peptidolytic degradation of natriuretic peptides
Natriuretic peptides (NP), like atrial- (ANP), B-type- (BNP)
and C-type natriuretic peptide (CNP), are cyclic peptide
hormones with relevance to cardiovascular, endocrine
and renal homeostasis. All three peptides contain an intact
17 amino acid disulfide-linked loop which is essential for
their biological activity. Two different mechanisms are discussed
as being responsible for the inactivation of the NP:
(i) binding to specific receptors (C-type-receptor) with
subsequent internalization and degradation,
(ii) degradation by extracellular peptidases.

The NEP is generally accepted to be the main enzyme
which initially catabolizes NP by cleavage within the loop
at the Cys-Phe bond.
In HPLC-monitored degradation studies we analyzed the
catabolism of mouse ANP and BNP by membranes of different
organs from NEP-deficient and wild-type mice. We
observed an unexpected result: Both NP peptides were
rapidly degraded by mouse lung and kidney membranes,
but in contrast to the generally published opinion, NEP was
only involved in the cleavage of ANP. A large percentage
of the ANP-degrading and the total BNP-degrading cannot
be ascribed to NEP. An interesting side result was the
up-regulation of the catabolic activity in NEP-deficient
membranes resulting in an unequivocally faster degradation
than in wild-type membranes. (Figure 1)
The next aim of the project will be the characterization of
this/these still unknown NP-degrading enzyme(s) and its
mode of action. A specific inhibition of these peptidases
would protect the NP. This would potentiate the cardioprotective
action of NPs and possibly enable new therapeutic
treatments.
In a second part of this project we want to find out the
molecular basis of the different degradation of NP by NEP.
In a first step, we proved the influence of the chemical
structure of NP on their catabolism by comparing NP of
different species with various structure modifications.
These degradation studies were performed with recombinant
NEP. The observed degradation rate was dependent
on the size of the N- and C-terminal tails of the NP. Only
human BNP was totally resistant in our test. These results
enabled a first model for the molecular interactions between
the catalytic center of NEP and NP (Figure 2).
To prove this model, we plan studies with NEP and NP
derivates from human BNP with (a) shorter terminal tails or
(b) replaced amino acids in the highly conserved innerloop
regions. This should help to detect why human BNP
is not a substrate for NEP. Moreover, modified peptidaseresistant
NP-analogues are interesting for potential therapeutic
use.
Group members
Dr. Winfried Krause**
Matthias Becker (Doctoral student)*
Xiaoou Sun (Doctoral student)
Kristin Pankow (Doctoral student)*
Bettina Kahlich (Technical assistance)
* part of period reported
** part-time
External funding
Deutsche Forschungsgemeinschaft
„Neuropeptidasen und Alkoholkonsum – Untersuchungen
an transgenen und knockout-Tieren“ (SI 483/3-1/2)
Wolf-Eberhard Siems
ASTA Medica
Wolf-Eberhard Siems
Strathmann AG
Wolf-Eberhard Siems
Selected publications (FMP authors in bold)
Gembardt F, Sterner-Kock A, Imboden H, Spalteholz M,
Reibitz F, Schultheiss H-P, Siems WE, Walther T (2005)
Organ specific distribution of ACE2 mRNA and correlating
peptidase activity in rodents. Peptides 26, 1270-1277
Heringer-Walther S, Moreira MC, Wessel N, Saliba JL,
Silvia-Barra J, Pena JL, Becker M, Siems WE, Schultheiss
HP, Walther T (2005) Brain natriuretic peptide predicts survival
in Chagas disease more effectively than atrial natriuretic
peptide. Heart 91, 385-387
Maul B, Walther T, Krause W, Pankow K, Becker M,
Gembardt F, Alenina N, Bader M, Siems WE (2005) Central
Angiotensin II Controls Alcohol Consumption via its AT1
Receptor. FASEB J 19:1474-1481
Walther T, Stepan H, Pankow K, Becker M, Schultheiss HP,
Siems WE (2004) Biochemical analysis of neutral endopeptidase
activity reveals independent catabolism of atrial
and brain natriuretic peptide. Biol Chem 385, 179-184
Walther T, Stepan H, Pankow K, Gembardt F, Faber R,
Schultheiss HP, Siems WE (2004) Relation of ANP and BNP
to their N-terminal fragments in fetal circulation: evidence
for enhanced neutral endopeptidase activity and resistance
of BNP to neutral endopeptidase in the fetus. BJOG
111, 452-455
Siems WE, Maul B, Wiesner B, Becker M, Walther T,
Rothe L, Winkler A (2003) Effects of kinins on mammalian
spermatozoa and the impact of peptidolytic enzymes.
Andrologia 35, 44-54
Collaborations
Thomas Walther, EMC Rotterdam, The Netherlands
Michael Bader, MDC Berlin
Doris Albrecht, Humboldt-Universität zu Berlin
Gisela Grecksch, University of Magdeburg
Sergej Danilov, University of Chicago, USA
Cellular Signalling/ Molecular Genetics
55

BIOPHYSICS
Group Leader: PD Dr. Peter Pohl
COMBINED TRANSPORT OF WATER AND
IONS THROUGH MEMBRANE CHANNELS
Life in all its diversity became possible due to the separation
of living entities from the lifeless and hostile environment
by means of membranes. It requires preservation of
selective transmembrane material and information
exchange. During evolution cells developed, the function
of which depends on the well-controlled interplay and
material exchange between different compartments. The
different transport functions are performed by a sophisticated
apparatus of membrane proteins, each of them is
fulfilling a very distinct function. Investigation of the latter
comprises the main interest of the biophysics group. We
aim to explore the molecular mechanisms of water, proton
and oxygen movement. Additional activities in the area
of applied biophysics are devoted to selected modifiers of
membrane permeability (e.g. ribosome inactivating proteins
and block copolymers). In this short report, we will
focus on water transport. The investigations are carried
out on different levels of organization: starting from proteins
reconstituted into planar and vesicular membranes,
followed by proteins overexpressed in different cell lines
and completed by proteins identified in primarily cultured
cell monolayers.
Water transport by aquaporins
Water transport is essential to all forms of life. Nevertheless,
the pathways taken by water across a membrane
barrier and the mechanism of solute-solvent coupling are
still not entirely resolved. Commonly, it is considered that
water may pass the lipid bilayer by diffusion. The extent to
which the lipid part of membranes contributes to water
transport varies considerably between different cells. Epithelial
cells, for example, have to maintain large chemical
and osmotic gradients. Consequently, the membrane
matrix has to be effectively impermeable to ions and most
other small molecules. Tightening of the lipid barrier requires
proteinaceous transport routes of water. Proteins specialized
on water transport are called aquaporins. We
have investigated the selectivity of several members of the
aquaporin family both in artificial model membranes and
in their cellular environment.
For example, in cooperation with the department “Molecular
Medicine” (Burkhard Wiesner, Dorothea Lorenz,
Walter Rosenthal) Dr. Tsunoda investigated the transport
mediated by aquaporin-1 (AQP1). It is a membrane channel
which allows rapid water movement driven by a trans-
membrane osmotic gradient. The protein was claimed to
have a secondary function as a cyclic-nucleotide gated
ion channel. However, upon reconstitution into planar
bilayers, the ion channel exhibited a tenfold lower single
channel conductance than in Xenopus oocytes and a hundredfold
lower open probability (

filter. These oscillations, where the chain of molecules
imbedded in the channel (the “liquid”) cooperatively exits
the channel leaving behind a near vacuum (the “vapor”),
so far have only been discovered in hydrophobic nanopores
by molecular dynamics simulations (Hummer, G.,
Rasaiah, J.C. & Noworyta, J.P. (2001) Nature 414, 188-190;
Beckstein, O. & Sansom, M.S.P. (2003) Proc. Natl. Acad.
Sci. USA. 100, 7063-7068).
Our results clearly show that in single file transport, water
molecules are more than just spacer molecules between
the ions. They are not only required for electrostatic reasons
but add important features to the overall transport
process. Whether the fast transport mode occurs in Na +,
Ca 2+, other K + channels or even in members of the aquaporin
family, which all realize single file transport, remains to
be elucidated, as well as the physiological importance of
this phenomenon. With respect to the high density of K +
channels in the nodes of Ranvier, for example, a contribution
to water homeostasis in neurons is likely.
Confinement of water into a very narrow geometry led to
profound changes of its physico-chemical properties. As a
result the mobility of water molecules inside the channel
and in the bulk differed by more than an order of magnitude.
Further investigation of the ultrafast diffusion will be
FIGURE 1
Fast osmotic water flow through the bacterial potassium
channel (KcsA) occurs only when all ions are rinsed out of
the filter. This situation is illustrated in the figure where all
binding sites are occupied by water molecules (red). The
potassium ion (yellow) has been placed into the cavity
which in contrast to the selectivity filter is wide enough to
allow different molecules to pass each other (Fig.3 from Biol.
Chem. 2004, 385: 921–926).
conducted in the frame of the priority program of the Deutsche
Forschungsgemeinschaft “Micro- and Nanofluidics”.
Group members
Dr. Sapar M. Saparov
Dr. Satoshi Tsunoda
Dr. Oxana O. Krylova
Dr. Valentina Margania**
Steffen Serowy (Doctoral student )*
Rustam Mollajew
Matthias Prigge (Student)*
Prof. Dr. Yuri Antonenko (Guest scientist)*
Dr. Valerij Sokolov (Guest scientist)*
Dr. Artem Ayuyan (Guest scientist)*
Christina de Souza (Guest scientist)*
Alexander Lentz (Guest scientist)*
Elena Sokolenko (Guest scientist)*
External funding
Deutsche Forschungsgemeinschaft
Heisenbergstipendium (Po533/7-1)
Peter Pohl
* part of period reported
** part-time
Cellular Signalling/ Molecular Genetics
57

Deutsche Forschungsgemeinschaft
“Single file water transport across peptidic nanopores”
(Po533/11-1)
Peter Pohl
Deutsche Forschungsgemeinschaft
„Molekulare Mechanismen des Wassertransports durch
Epithelzellmonoschichten“ (Po533/8-1, 8-2)
Peter Pohl
Deutsche Forschungsgemeinschaft
„Wechselwirkung zwischen ebener Bilipidmembran und
Photosensibilisator“ (Po533/4-3)
Peter Pohl
Deutsche Forschungsgemeinschaft
„Migration von Protonen an der Oberfläche ebener Bilipidmembranen“
(Po533/5-2, 5-3)
Peter Pohl
Deutsche Forschungsgemeinschaft
Kooperation mit der Lomonossov-Universität Moskau (436
RUS113/551)
Peter Pohl
Deutsche Forschungsgemeinschaft
Kooperation mit dem Frumkin-Institut für Elektrochemie
Moskau (436 RUS113/634)
Peter Pohl
Volkswagenstiftung
”Control of membrane permeability with novel types of
amphiphilic macromolecules” (I/77743)
Peter Pohl
Volkswagenstiftung
”Control of membrane permeability with novel types of
amphiphilic macromolecules” (I/80074)
Peter Pohl
Selected publications (FMP authors in bold)
Pohl P (2004) Combined transport of water and ions
through membrane channels. Biol Chem 385, 921-926
Saparov SM, Pohl P (2004) Beyond the diffusion limit:
Water flow through the empty bacterial potassium channel.
Proc Natl Acad Sci U. S. A 101, 4805-4809
Krylova OO, Pohl P (2004) Ionophoric activity of pluronic
block copolymers. Biochemistry 43, 3696-3703
Sun J, Pohl EE, Krylova OO, Krause E, Agapov II, Tonevitsky
AG, Pohl P (2004) Membrane destabilization by ricin.
Eur Biophys J 33, 572-579
Tsunoda SP, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin-1, Nothing but a Water Channel. J Biol
Chem 279, 11364-11367
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W, Pohl
P, Maric K (2003) Cyclic AMP is sufficient for triggering the
exocytic recruitment of aquaporin-2 in renal epithelial
cells. EMBO Rep 4, 88-93
Serowy S, Saparov SM, Antonenko YN, Kozlovsky W,
Hagen V, Pohl P (2003) Structural Proton Diffusion along
Lipid Bilayers. Biophys J 84, 1031-1037
Collaborations
- Proton migration (DFG Po 533/5-1, 2, 3; 436 RUS113/551):
Prof. Yuri N. Antonenko, Lomonossov University Moscow,
Belozersky Laboratory
- Photosensibilisator (DFG Po 533/4-3; 436 RUS113/634):
Dr. Valerij Sokolov, Russian Academy of Science, Frumkin
Institute of Electrochemistry
- Blockcopolymers and multidrug resistance (VW): Prof.
J. Kressler, Martin-Luther-Universität Halle; Prof. Dr.
Frey, University of Mainz

CYTOKINE SIGNALING
Group Leader: Dr. Klaus-Peter Knobeloch
FUNCTIONAL ANALYSIS OF POSTTRANSLA-
TIONAL PROTEIN MODIFICATION BY UBI-
QUITIN AND UBIQUITIN LIKE MOLECULES
USING GENE TARGETING IN THE MOUSE
Covalent attachment of Ubiquitin and Ubiquitin like (UBL)
molecules serves as an important regulatory mechanism
controlling a pleiotropy of biological processes including
embryogenesis, cell cycle, growth control, and immune
response. Beside serving as a marker that targets proteins
for proteasomal degradation Ubiquitin conjugation has
been demonstrated to be involved in the regulation of a
wide range of molecular mechanisms like protein localization,
receptor degradation, transcriptional control and
interaction of proteins.
Thus distinct components of the Ub and UBL conjugation
and deconjugation pathway might serve as novel targets
for drug intervention.
In our group targeted mutagenesis in the mouse is used
to address the biological relevance and function directly
in the context of the whole organism. Current research is
focused on ISG15, an interferon stimulated ubiquitin like
molecule and UBPy, a deubiquitinating enzyme.
ISG15 is one of the most strongly induced genes upon
interferon treatment and like SUMO or NEDD8 belongs to
the family of ubiquitin-like modifiers. ISG15 can be conjugated
to distinct proteins and has been implicated in a
variety of biological activities which encompass antiviral
defence, immune responses, and pregnancy. Mice lackin
UBP43 (USP18), the ISG15 deconjugating enzyme which
belongs to the family of deubiquitinating enzymes, develop
a severe phenotype with brain injuries and lethal
hypersensitivity to Poly(I:C). It was reported that an augmented
conjugation of ISG15 in the absence of UBP43
induces prolonged STAT1 phosphorylation and that the
ISG15 conjugation plays an important role in the regulation
of JAK/STAT and interferon signaling (Malakhova OA et
al. 2003, Genes & Development 17, 455-460). It was also
published that UBP43-/-mice are more resistant against
infection with lymphocytic choriomeningitis virus (LCMV)
and vesicular stomatitis virus (VSV).
To directly assess the functional role of ISG15, we generated
mice lacking ISG15 via gene targeting in embryonic
stem cells. ISG15-/- mice display no obvious abnormalities,
and lack of ISG15 did not affect the development and composition
of the main cellular compartments of the immune
system. In contrast to UBP43-/- mice, the interferon-induced
antiviral state and immune responses directed against
VSV and LCMV were not significantly altered in the absence
of ISG15. Furthermore, interferon or endotoxin induced
STAT1 tyrosine-phosphorylation, as well as expression of
typical STAT1 target genes remained unaffected by the
lack of ISG15. Thus the role of ISG15 in the pathology of the
phenotype of UBP43-/- and the specificity of UBP43 needs
to be reassessed. Therefore, we generated mice deficient
for both molecules in an attempt to rescue phenotypic
alterations of UBP43-/- mice, providing they are caused by
an enhanced ISG15 conjugation. This approach is also
suitable to test whether UBP43 possesses additional
functions beside acting as an ISG15 deconjugating enzyme.
These double knockout mice are currently under analysis.
We were also able to clone a new mouse gene, which we
named UBL14, that encodes a protein with 95% amino acid
homology to ISG15. To elucidate the role of this molecule
in vivo, we also used a loss of function approach and
generated mice deficient for UBL14, as well as for both,
ISG15 and UBL14.
Conditional inactivation of the ubiquitin-specific
protease UBPy/USP8
Ubiquitination of proteins is now recognized to target proteins
for degradation by the proteasome and for internalization
into the lysosomal system, as well as to modify
functions of some target proteins. Although much progress
has been made in characterizing enzymes that link ubiquitin
to proteins, the understanding of deubiquitinating enzymes
is just beginning to evolve. The importance of deubiquitinating
enzymes has been demonstrated by the
observation that cellular key molecules like p53 or NFkappa
B are controlled by ubiquitin isopeptidases (Li et al
2002, Trompouki et al 2003). However, so far there are no
reports challenging the function of deubiquitinating proteins
in the context of the whole organism.
UBPy (USP8) is an ubiquitin isopeptidase that is upregulated
upon serum induction and was shown to bind to SH3
domains of certain target proteins via an unusual recognition
motif. From patients with a myeloproliferative disorder
a fusion product of the p85beta subunit of phosphatidylinositol-3-kinase
and UBPy could be isolated. In an initial
approach we have generated mice lacking UBPy. As these
mice die early in embryogenesis we generated mice that
allow conditional inactivation of the gene using the creloxP
system. By mating the mice to Mx-cre we have inactivated
the gene in adult mice and are analyzing the
phenotypic alterations, and molecular mechanisms that
are affected by the lack of UBPy. In parallel, we genera-
Cellular Signalling/ Molecular Genetics
59

FIGURE 1
ISG 15 conjugation cycle
ted mice lacking UBPy in specific hematopoietic cell lineages
like T-cells and granulocytes/macrophages in order to
determine the functional role of UBPy in these cell lineages.
Group members
Sandra Niendorf (Doctoral student)
Agnes Kisser (Doctoral student)
Markus Wietstruck (Technical assistance)
Anna Osiak (Doctoral student)*
Liane Boldt (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Herstellung und Haltung genetisch veränderter Mäuse“
(TP Z3 im Sonderforschungsbereich 366 „Zelluläre Signalerkennung
und Umsetzung“)
Ivan Horak
Deutsche Forschungsgemeinschaft
„Untersuchungen zur Funktion des Interferon-stimulierten
Genes 15 (ISG15).“ (KN 590/1-1)
Klaus-Peter Knobeloch
* part of period reported
FIGURE 2
Editing functions of deubiquitinating enzymes. Deubiquitinating enzymes
may negatively regulate proteolysis or other signaling functions
of ubiquitination such as internalization or altered protein function by
removing the ubiquitin chain from the target proteins.
Selected publications (FMP authors in bold)
Osiak A, Utermohlen O, Niendorf S, Horak I, Knobeloch KP.
(2005) ISG15, an interferon-stimulated ubiquitin-like protein,
is not essential for STAT1 signaling and responses
against vesicular stomatitis and lymphocytic choriomeningitis
virus.
Mol Cell Biol., 6338-6345
Van Spriel AB, Puls KL, Sofi M, Pouniotis D, Hochrein H,
Orinska Z, Knobeloch K-P, Plebanski M, Wright MD (2004)
A regulatory role for CD37 in T-Cell Proliferation. J Immunol
172, 2953-2961
Rosenbauer F, Wagner K, Zhang P, Knobeloch KP, Iwama
A, Tenen DG (2004) pDP4, a novel glycoprotein secreted by
mature granulocytes, is regulated by transcription factor
PU.1. Blood 103, 4294-4301
Schuh K, Cartwright EJ, Jankevics E, Bundschu K, Liebermann
J, Williams JC, Armesilla AL, Emerson M, Oceandy
D, Knobeloch KP, Neyses L (2004) Plasma membrane Ca2+
ATPase 4 is required for sperm motility and male fertility. J
Biol Chem 279, 28220-28226

Huebner A, Kaindl AM, Knobeloch KP, Petzold H, Mann P,
Koehler K (2004) The triple A syndrome is due to mutations
in ALADIN, a novel member of the nuclear pore complex.
Endorcr Res 30, 891-899
Collaborations
International
Herbert W. Virgin IV, Department of Pathology
Washington University School of Medicine
Prof. Dr. Robert Krug, Institute for Cellular and Molecular
Biology
University of Texas at Austin
National
Prof. Dr. med. Oliver Liesenfeld, Institut für Mikrobiologie
und Hygiene
Charité – Universitätsmedizin Berlin
PD Dr. Marco Prinz, Abt. Neuropathologie
Georg-August-Universität Göttingen
Dr. Olaf Utermöhlen, Institut für Medizin. Mikrobiologie,
Immunologie und Hygiene
Universität zu Köln
Cellular Signalling/ Molecular Genetics
61

MOLECULAR MYELOPOIESIS
Group Leader: Dr. Dirk Carstanjen
ICSBP – A CRITICAL TRANSCRIPTION
FACTOR FOR MYELOPOIESIS
The research aim of our group is to elucidate the role of
the transcription factor ICSBP (interferon consensus
sequence binding protein) within hematopoietic cell development.
This hematopoietic specific transcription factor
is a major element regulating transcriptional control of
myelopoiesis. Mice deficient in this transcription factor
develop a proliferative syndrome characterized by the
expansion and accumulation of immature myeloid progenitor
cells within the bone marrow and spleen. Furthermore,
it has been shown by our group that ICSBP influences
lineage choice of granulocyte-monocyte progenitor
cells (GMP) leading to an imbalance in development and
maturation of progenitor cells in favor of granulocytes.
Genetic profile of myeloid development
This asymmetric lineage choice is at least partially due to
a reduced response to a macrophage specific cytokine
(macrophage colony stimulating factor: M-CSF). Our group
has recently shown that M-CSF signal transduction in
macrophages is altered due to altered protease expression
in these cells and enhanced proteasomal degradation
leading to shortened signaling. In contrast, the reason for
accumulation of immature progenitor cells and preferential
production of granulocytes is further obscure. In an
approach to decipher the function of ICSBP in immature
progenitor cells as well as to define the expression profile
of granulocyte versus erthrocyte precursors, we sorted
GMP and erythrocyte progenitor (EP) cells by flow cytometry
in collaboration with H.R. Rodewald, Ulm. Global
gene expression profiling was performed in these highly
purified cells and analyzed using Affymetrix technology.
The data sets of this analysis have been published via the
GEO database. These data now allow for the first time to
study systematically the transcriptional profile of these
functional distinct cell populations. As a first approach, we
identified the global pattern of transcription factors specifically
expressed in either lineage. While this analysis confirmed
the exclusive or highly preferential expression of
several transcription factors with pivotal importance for
erythroid development (e.g. erythroid Krüppel-like factor,
GATA-1, Friend of GATA-1 (FOG1)), we also identified several
transcription factors specifically expressed in myeloid
progenitors. Several of those are known: Cebpα, Cebpβ,
Gfi1, Fli1, PU.1, and Meis1 to name but a few, others have
not yet been implicated in myeloid development, e.g.
Mef2c, KLF4, and SAT1b. Importantly, ICSBP has been
shown to be specifically expressed by GMP, and the RNA
expression level was within the group of the five transcription
factors the most prominently expressed transcription
factors in these cells, in concert with prominent proteins
as PU.1, and Cebpα. These important findings further confirms
the pivotal role of ICSBP for myeloid development
and will now allow to study systematically the analysis of
transcription factors regulating myeloid and erythroid cell
development using genetic approaches e.g., mouse deletion-mutants,
RNA interference or retroviral overexpression.
These studies are currently being undertaken.
Dissecting the transcriptional network orchestrated
by ICSBP
We further analyzed the global gene expression profile in
the GMP of the ICSBP mouse deletion mutant. This analysis
revealed insight into the developmental program induced
by the loss of ICSBP within myeloid progenitor cells.
We found several major myeloid transcription factors
downregulated in the GMP of ICSBP-deleted mice. We are
currently studying the contribution of these transcription
factors to the myeloproliferative syndrome of these mice
using mouse deletion mutants and retroviral overexpression
in bone marrow derived stem and progenitor cells.
Furthermore, we are studying potential direct transcriptional
regulation of ICSBP on genomic target sequences. We
therefore cloned several murine genomic sequences with
known or potential regulatory functions for the genes we
found differentially expressed in GMPs of ICSBP-deleted
mice. Luciferase promoter studies are currently being
undertaken to identify the impact on ICSBP in direct or
combinatorial gene expression regulation with other prominent
myeloid transcription factors. Furthermore, many
of the genes overexpressed in ICSBP-deleted mice are
genes preferentially expressed in mature granulocytic and
monocytic cells. Interestingly, at the GMP stage, there was
no clear preferential expression pattern of granulocytic
genes implicating that branching between monocytic and
granulocytic development occurs beyond the GMP stage
and additional extrinsic factors, e.g. cytokine signaling, are
responsible for the preferential generation of granulocytes
versus macrophages in the ICSBP deleted mouse, a
hypothesis consistent with our previous findings showing
abnormal M-CSF signaling in macrophages derived from
the ICSBP deleted mouse.

c-Kit
IL-3Rα
Group members
Maja Djurica (Doctoral student)*
Jessica Königsmann (Doctoral student)*
Axel Kallies (Doctoral student)*
Joanna Selfe (Doctoral student)*
Melanie Benedict (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Myeloproliferatives Syndrom bei ICSBP-defizienten Mäusen:
Identifizierung neuer potentieller onkotherapeutischer
Targetstrukturen“ (TP G1 im Sonderforschungsbereich
506 „Onkotherapeutische Nukleinsäuren“)
I. Horak
Deutsche Forschungsgemeinschaft
„Gene in Entwicklung und Funktion von myeloiden Zellen“
HO 493/12-1 und 12-2
Ivan Horak
* part of period reported
GMP
GMP
FIGURE 1
Bone marrow cells from ICSBP wt (upper) and deleted
(lower part) were depleted of mature cells expressing marker
for granulocytes, monocytes, T-, B-cells and erythrocytes.
Those so-called lineage negative cells were further
stained with antibodies against c-kit (stem cell factor receptor)
as well as interleukin-3 receptor. c-kit high, Il-3 receptor
positive cells contain so called granulocyte-monocyte
progenitor cells (GMP), megakaryocyte progenitor cells
(MP), and common myeloid progenitor cells (CMP). Those
populations can be identified through the use of antibodies
against FcγR I and CD41. The GMP population contains only
cells that give rise to monocytes/macrophages and granulocytes.
This population was sorted to purity, RNA isolated,
and global gene expression profiling was performed using
Affymetrix technology.
Deutsche Forschungsgemeinschaft
„Die Rolle des Adaptorproteins Disabled-2 bei Zytokinsignalvermittlung
in hämatopoetischen Zellen“ (CA 306/1-1)
Dirk Carstanjen
Selected publications (FMP authors in bold)
Zatóvicova M, Tarabkova K, Svastova E, Gibadulinova A,
Mucha V, Jakubickova L, Biesova Z, Rafajova M, Ortova Gut
M, Parkkila S, Parkkila AK, Waheed A, Sly WS, Horak I,
Pastorek J, Pastorekova S (2003) Monoclonal antibodies
generated in carbonic anhydrase IX-deficient mice recognize
different domains of tumour associated hypoxia-induced
carbonic anhydrase IX. J Immunol Methods 282, 117-134
Barmeyer C, Harren M, Schmitz H, Heinzel-Pleines U,
Mankertz J, Seidler U, Horak I, Wiedenmann B, Fromm M,
Schulzke JD (2004) Mechanisms of diarrhea in the interleukin-2-deficient
mouse model of colonic inflammation.
Am J Physiol Gastrointest Liver Physiol 286, G244-G252
Terszowski G, Waskow C, Conradt P, Lenze D, Koenigsmann
J, Carstanjen D, Horak I, Rodewald HR (2004) Prospective
isolation and global gene expression analysis of
the colony-forming unit-erythrocyte (CFU-E). Blood 105,
1937-1945
Cellular Signalling/ Molecular Genetics
63

MOUSE MODELS
Group Leader: Dr. Rüdiger Pankow
BCL-2 ASSOCIATED TRANSCRIPTION
FACTOR AND FMS INTERACTING PROTEIN
Btf (Bcl-2 associated transcription factor) and FMIP (fms
interacting protein) are two genes which have been implicated
in cellular regulatory processes. Both are intracellular
proteins with subcellular distributions in the cytoplasm
and the nucleus. We have raised antibodies against
Btf and found its expression in a broad spectrum of tissues
and organs including those of the hematopoietic
system. Its highest expression, however, is found in brain
regions of young developing mice. This expression declines
with age, with Btf being detectable in only a few
neural cells in adulthood.
Due to its association with Bcl-2, Btf was thought to be a
transcriptional repressor, with an apoptotic function. Since
no rigorous evidence for a role of Btf in apoptosis was
found, its biological function remains unknown. To generate
mice deficient for Btf we inserted a marker cassette
containing the β-galactosidase gene into the btf gene. The
inserted β-galactosidase gene is under the control of the
endogenous btf promoter which allows the detection of
btf gene expression. Mice, homozygous for this mutation,
lack all wild type btf transcripts and are deficient in
Btf protein. These mice display a severe phenotype resulting
in premature death. Molecular mechanisms behind
the phenotypic changes are being analyzed. To assess the
role of Btf in the hematopoietic system we reconstituted
lethally irradiated wild type mice with fetal liver or bone
marrow cells from Btf deficient mice. These mice reconstituted
a Btf -/- hematopoietic system including all resulting
cell lineages but do not develop any of the phenotypic
signs characteristic of Btf deficient mice. We could therefore
rule out the influences of the hematopoietic system
and conclude that other factors provide the major cause
for the phenotype.
For FMIP, an interacting partner of the activated M-CSF
receptor, a possible role in myeloid differentiation was
suggested. By means of targeted gene disruption, we
generated a mouse model with a null mutation of the fmip
gene. Our studies revealed, however, that embryos lacking
Fmip die early during embryogenesis. Thus, to allow studies
of Fmip in physiological processes in the mouse, we
have generated FMIP conditional knockout mice using the
cre/loxP system. By intercrossing these mice with Credeleter
mouse strains we have obtained Fmip floxed mice
that allow organ or cell lineage specific deletion of Fmip
and have used these models to study the function of FMIP
and in particular, its relevance in myeloid development.
Group members
Dr. Rosel Blasig
Kerstin Bohne (Technical assistance)
Janet Klemm (Technical assistance)
Mina Thakur (Technical assistance)*
* part of period reported

CELLULAR SIGNAL PROCESSING
Group Leader: Dr. Uwe Vinkemeier
KINETIC CONTROL OF GENE TRANSCRIPTION
We are studying the effects on gene transcription of a
group of extracellular signaling molecules termed cytokines,
more than a hundred of which are presently known.
Cytokines are secreted small proteins that fulfill crucial
roles in cell differentiation and innate immunity. Best
known are the interleukins and erythropoietine (Epo) for
their role in immune cell differentiation, inflammation and
lactation. Other cytokines such as LIF and Oncostatin M
control stem cell development and cell growth, while the
interferons are known to cause growth arrest and protect
cells against viral and microbial attack. Not surprisingly,
cytokines are highly relevant for clinical applications.
Several cell-activating cytokines are being tested in clinical
trials, and interleukin-2 was the first drug approved for
the treatment of tumors. Today, treatment with interferons
is the standard therapy for several severe viral infections
(e.g. hepatitis) as well as for some tumors (e.g. skin cancer).
However, overexpression of cytokines may also
cause diseases such as rheumatoid athritis. In those
situations, inhibition of cytokine action is a therapeutic
goal. As the alteration of gene transcription is the basis of
cytokine action, we need to explore the intracellular
events that are triggered by cytokines in order to understand
their effects in the normal and diseased state.
The Janus kinase (JAK) / signal transducer and activator
of transcription (STAT) pathways, first identified in the
interferon systems, are responsive to a wide range of cytokines
and growth factors. The STAT proteins receive cytokine
signals at intracellular receptor chains in the cytoplasm
and carry them into the nucleus, where they then
act as transcription factors. Thus, these proteins need to
cross the nuclear envelope to functionally link the cell
membrane with the promoters of cytokine-responsive
genes. Movement of STATs in either compartment is diffusion
controlled and not directed along permanent structures.
However, passage through the nuclear gateways,
named nuclear pore complexes (NPC), provides a formidable
diffusion barrier to proteins the size of STATs (> 85
kD for the monomer), as only ions and small molecules not
exceeding 40 kD can freely enter the cell nucleus. The
analysis of their nucleocytoplasmic translocation has to
consider that the STAT proteins exist in two different states
in terms of signaling: before the stimulation of cells with
cytokines the STAT molecule exists in a non-tyrosine phosphorylated
state. Stimulation with cytokines increases the
activity of receptor-associated JAK kinases, leading to the
formation of tyrosine-phosphorylated STATs, which
instantly assemble into homo- or heterodimers via canonical
phosphotyrosine-SH2 domain interactions. Tyrosine
phosphorylation is often described as STAT activation
since only the dimer is a high-affinity DNA-binding protein
required for the induction of cytokine-responsive genes.
This is achieved by directly targeting cognate recognition
elements named gamma activated sites (GAS).
Carrier-dependent as well as carrier-independent modes
of translocation are known to exist for the passage of proteins
through the nuclear pore. The nuclear pore complex
is a multi-protein structure that creates an aqueous channel
spanning the double membrane of the nuclear envelope.
This structure with an estimated molecular mass of
125 MDa in mammalian cells is composed of several
copies of about 30 different proteins collectively called
nucleoporins (Nups), many of which contain multiple phenylalanine-glycine
(FG) repeats that are interspersed with
polar residues of varying number. Although the molecular
details that allow passage through the pore remain to be
elucidated, it is widely accepted that permeating molecules
need to overcome the hydrophobic repulsion that is
exerted by the FG repeat-rich interior of the nuclear pore.
Biochemically carrier-free and carrier-dependent nucleocytoplasmic
translocation are related. Docking to the
nuclear pore is diffusion controlled for the two pathways,
and in both cases the actual translocation process appears
to occur independent of metabolic energy via identical
interactions with channel components. Proteins that
contain regions that enable them to directly engage in productive
interactions with the nucleoporins are capable of
carrier-independent nucleocytoplasmic transport, whereas
the remaining proteins (also called cargo proteins)
need to associate with transport factors, which act as
chaperones during passage through the pore. The vast
majority of transport factors belongs to the karyopherin
superfamily of proteins, which mediate either import or
export from the nucleus. Recognition of cargo proteins by
the transport factors requires the presence of loosely conserved
stretches of amino acids on the cargo surface, termed
nuclear localization -NLS- or nuclear export -NESsignals,
respectively. The stability of cargo/karyopherin
complexes is determined by the small GTPase Ran, as
RanGTP disrupts importin/cargo complexes but stabilizes
the formation of exportin/cargo-complexes. Thus, the
RanGDP/RanGTP gradient that forms through the asymmetric
distribution of nucleotide exchange factors across the
nuclear membrane discriminates cytosol from nucleoplasm
and hence confers directionality to the transport
process.
Cellular Signalling/ Molecular Genetics
65

Our lab discovered that the STATs make use of both of
these translocation mechanisms. Due to their constant
carrier-independent and -dependent nucleocytoplasmic
cycling the STATs are distributed throughout the cell, all
the time. Deviations from an even pancellular distribution
result from modulated nuclear export. This is caused either
by enhancing the translocation rate or by nuclear
retention due to a co-factor-independent mechanism. The
recognition that the dephosphorylation reaction is under
kinetic control of DNA binding provided a strikingly simple
example of a self-controlling mechanism that integrates
central elements of cytokine-dependent gene regulation,
namely receptor monitoring, promoter occupancy, and
transcription factor activity. It is important to point out that
nuclear accumulation is not a mechanism sui generis, but
merely reflects the formation of a conformation that is resistant
to dephosphorylation. For this reason nuclear accumulation
is not suited as a diagnostic marker of transcriptional
activity. In contrast, the translocation rate of STAT
transcription factors critically determines the outcome of
cytokine signaling and hence constitutes a potential specificity
determinant. The molecular mechanisms that give
rise to flux modulation, either physiologically or in pathological
situations such as microbial infections, and how
this affects both duration and strength of cytokine signals,
as well as the overall sensitivity and latency of the system
will be the task of future research. A further important area
of our research is post-translational modifications, with
focus on arginine methylation and sumoylation.
In the last years our lab has made a number of exciting and
unexpected discoveries, which revealed that the STATs
are an intriguing model system for studying the intracellular
dynamics of a signal transducer. In addition, it is increasingly
being appreciated that STAT nucleocytoplasmic
cycling also constitutes an important area for microbial
intervention. Currently, we are generating mice that
express STAT mutants. This will allow us to study the
effects of defective nucleocytoplasmic shuttling on development
and disease. These approaches are complemented
by chemical and structural biology to explore the
possibilities of rational intervention.
Group members
Prof. Dr. mult. Thomas Meyer
Dr. Andreas Begitt
Dr. Regis Cartier
Dr. Ying Shan**
Dr. Andreas Marg*
Torsten Meissner (Doctoral student)*
Inga Lödige (Doctoral student)
Nicola Venta (Doctoral student)*
* part of period reported
** part-time
Mandy Kummerow (Technical assistance)
Stephanie Meyer (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Konstitutiver nucleocytoplanmatischer Transport von
STAT1“ (VI 218/2)
Uwe Vinkemeier Deutsche Forschungsgemeinschaft
Deutsche Forschungsgemeinschaft
„Einfluss der Tyrosin-Dephosphorylierung auf die Zielgenfindung
von STAT1“ (VI 218/3)
Uwe Vinkemeier
Deutsche Forschungsgemeinschaft
„Analyse einer konservativen Protein-Interaktionsdomäne“
(VI 218/4)
Uwe Vinkemeier
Bundesministerium für Bildung und Forschung
„Molekulare Grundlagen der zellulären Signalverarbeitung“
(Sieger im Nachwuchsgruppenwettbewerb „Bio-
Future“)
Uwe Vinkemeier
European Molecular Biology Organization
“Regulation of Transcription” (EMBO-Young-Investigator
Award 2002)
Uwe Vinkemeier
Selected publications (FMP authors in bold)
Chen X, Bhandari R, Vinkemeier U, Van Den Akker F, Darnell
JE Jr, Kuriyan J (2003) A reinterpretation of the dimerization
interface of the N-terminal domains of STATs. Protein
Sci 142, 361-365
Meyer T, Vinkemeier U, Meyer U (2003) Medizinische
Implikationen pharmakogenomischer Behandlungsstrategien.
Ethik in der Medizin 12, 207-209
Meyer T, Vinkemeier U, Meyer U (2003) Evidence-based
medicine – Was geht verloren? Ethik in der Medizin 14,
3-10
Meyer T, Marg A, Lemke P, Wiesner B, Vinkemeier U
(2003) DNA binding controls inactivation and nuclear
accumulation of the transcription factor Stat1. Genes Dev
17, 1992-2005

Marg A, Shan Y, Meyer T, Meissner T, Brandenburg M,
Vinkemeier U (2004) Nucleocytoplasmic shuttling by
nucleoporins Nup153 and Nup214 and CRM1-dependent
nuclear export control the subcellular distribution of latent
Stat1. J Cell Biol 165, 823-833
Meissner T, Krause E, Vinkemeier U (2004) Ratjadone and
leptomycin B block CRM1-dependent nuclear export by
identical mechanisms. FEBS Lett 576, 27-30
Meissner T, Krause E, Lödige I, Vinkemeier U (2004)
Arginine Methylation of STAT1: A Reassessment. Cell 119,
587-589
Meyer T, Hendry L, Begitt A, John S, Vinkemeier U (2004)
A single residue modulates tyrosine dephosphorylation,
oligomerization, and nuclear accumulation of stat transcription
factors. J Biol Chem 279, 18998-19007
Meyer T, Vinkemeier U (2004) Nucleocytoplasmic shuttling
of STAT transcription factors. Eur J Biochem 271, 4606-
4612
Vinkemeier U (2004) Getting the message across STAT!
Design principles of a molecular signaling circuit. J Cell
Biol 167, 197-201
Collaborations
Susan John, Kings’s Kollege London, Department of Immunobiology
Ilian Jelesarov and H. R. Bosshard, Universität Zürich,
Institut für Biochemie
Gino Cingolani, Stony Brook University, USA
Ulrich Kubischeck, Friedrich-Wilhelm-Universität Bonn,
Institut für Physikalische und Theoretische Chemie
Markus Czub, Canadian Science Center for Human and
Animal Health, Winnipeg, Canada
Thomas Höfer and Reinhard Heinrich, Institut für Theoretische
Biophysik, Humboldt-Universität zu Berlin
Cellular Signalling/ Molecular Genetics
67

CHEMICAL BIOLOGY

SECTION CHEMICAL BIOLOGY
Prof. Dr. Michael Bienert
Department Head: Peptide Chemistry
(Secretary: Marianne Dreissigacker)
Hartmut Berger
Michael Beyermann
Margitta Dathe
Volker Hagen
Eberhard Krause
Jens-Peter von Kries
Johannes Oehlke
Jörg Rademann
Although in the last decade our knowledge about genome
sequences (genomics) and the occurrence of cellular proteins
(proteomics) has increased dramatically, this progress
is often not sufficient to understand the function and
the significance of biomacromolecules in cellular systems
or in organs of different organisms. All proteins evoke their
function by specific interactions with other proteins or
constituents of membranes and the cytoskeleton, and dee-
INTRODUCTION
BEREICH CHEMISCHE BIOLOGIE
Prof. Dr. Michael Bienert
Abteilungsleiter: Peptidchemie
(Sekretariat: Marianne Dreissigacker)
Hartmut Berger
Michael Beyermann
Margitta Dathe
Volker Hagen
Eberhard Krause
Jens-Peter von Kries
Johannes Oehlke
Jörg Rademann
Obwohl unser Wissen über Gensequenzen (Genomics)
und über das Auftreten zellulärer Proteine (Proteomics)
dramatisch angewachsen ist, reicht der Fortschritt oft
noch nicht aus, um die Funktion und Bedeutung von biologischen
Makromolekülen in zellulären Systemen oder in
den Organen der verschiedenen Organismen zu verstehen.
Alle Proteine funktionieren, indem sie spezifische
Interaktionen mit anderen Proteinen oder mit Bestandteilen
der Membranen und des Zytoskeletts eingehen, und
tiefergehende Einsichten auf molekularer und atomarer
per insights at a molecular and atomic level are required
in order to unravel the subtle modes of protein-protein
communication. The analysis of interactions between chemical
moieties of proteins, and the identification of protein
substructures, affected by drug-like compounds, as well
as the design and the synthesis of compounds with modified
recognition characteristics, demand chemical thinking
and chemical methodologies. This makes clear that
chemistry has become a more central science in our
efforts to understand the molecular basis of life. This dramatic
trend has led to the creation of the term “Chemical
Biology”, defining all kinds of chemical research focused
on the unravelling of biological phenomena. Naturally,
there is an overlap with other areas of chemistry, such as
“Medicinal Chemistry” and “Bioorganic Chemistry”. The
interaction with a powerful Structural Biology and the
involvement of biochemical, biophysical and analytical
methodologies are necessities for the further successful
development of the novel science “Chemical Biology”.
In the context of Chemical Biology, chemical and biological
research in the FMP are traditionally excellently interconnected.
Research efforts of the Department of Peptide
Ebene sind nötig, um die Feinstruktur der Protein-Protein-
Kommunikation zu erfassen. Sowohl die Analyse der
Wechselwirkungen zwischen chemischen Resten an Proteinen
und Identifizierung von Proteinsubstrukturen, die
durch Wirkstoffe beeinflussbar sind, als auch das Design
und die Synthese von Verbindungen mit veränderter
Erkennungscharakteristik erfordern chemisches Denken
und chemische Methodologie. Dies macht klar, dass die
Chemie für unseren Bemühungen, die molekulare Basis
des Lebens zu verstehen, zu einer zentralen Wissenschaft
geworden ist. Dieser dramatische Trend hat zur Entstehung
des Begriffs „Chemische Biologie“ geführt, der
alle Arten chemischer Forschung beschreibt, die auf die
Untersuchung biologischer Phänomene gerichtet sind.
Natürlich gibt es Überlappungen mit anderen Gebieten der
Chemie, wie zum Beispiel der Medizinischen und der Bioorganischen
Chemie. Datenaustausch mit einer starken
Strukturbiologie und die Einbeziehung biochemischer, biophysikalischer
und analytischer Methoden sind eine Notwendigkeit
für eine weitere erfolgreiche Entwicklung der
jungen Wissenschaft Chemische Biologie.
Im Kontext der Chemischen Biologie ist die chemische und
biologische Forschung am FMP traditionell exzellent vernetzt.
Forschungsarbeiten der Abteilung Peptidchemie

Chemistry and Biochemistry are focused on the molecular
mechanisms of peptide-protein (Peptide Synthesis Group,
Peptide Biochemistry Group) and peptide-lipid interactions
(Peptide-Lipid Interaction Group), leading to a better
understanding of the action of biologically active peptides
on their respective targets. The study of protein-protein
interactions in the cell requires the improvement of the
cellular uptake of peptides and other low-molecular compounds
(Peptide-Lipid Interaction Group). The function of
proteins is often regulated by distinct chemical, posttranslational
modification, which can be analyzed by
newly developed mass spectrometric procedures (Mass
Spectrometric Group). In addition, the department is involved
in methodological studies in order to improve the
chemical synthesis of small-sized proteins, as well as to
synthesize fluorescently labeled peptide analogues for
localization and interaction studies.
Very useful tools for the study of intracellular biological
processes, “caged compounds”, are synthesized and
characterized in the Synthetic Organic Biochemistry
Group. Caged compounds are photolabile, inactive derivatives
of biologically active substances, from which the
und Biochemie zielen auf die molekularen Mechanismen
von Peptide-Protein- (Arbeitsgruppe Peptidsynthese;
Peptidbiochemie) und Peptid-Lipid-Interaktionen (Arbeitsgruppe
Peptid-Lipid Interaktion) und verhelfen zu einem
besseren Verständnis der Wirkung biologisch aktiver Peptide
auf ihre jeweiligen Zielstrukturen. Die Untersuchung
von Protein-Protein Interaktionen in der Zelle erfordert
eine verbesserte Aufnahme der Peptide und anderer kleiner
Moleküle (Arbeitsgruppe Peptid-Lipid Interaktion). Die
Funktion von Proteinen ist oftmals durch bestimmte chemische
posttranslationale Modifikationen reguliert, die
mittels neuartiger massenspektrometrischer Verfahren
analysiert werden können (Arbeitsgruppe Massenspektrometrie).
Darüber hinaus ist die Abteilung an methodologischen
Studien beteiligt, die die chemische Synthese
kleiner Proteinmoleküle verbessern und fluoreszenzmarkierter
Peptidanaloga für Lokalisierungs- und Interaktionsstudien
gewährleisten sollen.
Äußerst nützliche Werkzeuge für die Untersuchung intrazellulärer
biologischer Prozesse, „caged compounds”,
werden in der Arbeitsgruppe Synthetische Organische
Biochemie synthetisiert und charakterisiert. Caged compounds
sind photolabile, inaktive Derivate biologisch
aktiver Substanzen, die aktive Biomoleküle, z. B. einen
active biomolecule, e. g. a transmitter, is rapidly liberated
by UV light, thus allowing the analysis of fast biological
processes in cells.
In 2004, Chemical Biology at the FMP was considerably
strengthened by the installation of the Medicinal
Chemistry Group. A central aim of this group consists in
the creation and development of synthetic methods for the
combinatorial organic chemistry and solid phase organic
synthesis. Focused libraries of potential ligands are generated
in order to find small organic molecules which bind
specifically to selected target proteins, thus assisting in
the analysis of functions and interactions of the proteins.
Naturally, such small molecules could also serve as lead
structures for the design of drug candidates. For analyzing
the biological activity of small-molecule libraries, a
Screening Unit is now established, developing concepts
for the handling of libraries and providing the technical
equipment for testing hundreds and thousands of chemical
compounds.
Transmitter, auf Einwirkung von ultraviolettem Licht hin
freisetzen und so die Analyse schneller biologischer Prozesse
in Zellen ermöglichen.
Im Jahr 2004 wurde die Chemische Biologie am FMP durch
die Einrichtung der Arbeitsgruppe Medizinische Chemie
beträchtlich gestärkt. Ein zentrales Ziel der Gruppe ist die
Entwicklung und Etablierung von Methoden für die kombinatorische
organische Chemie und Festphasensynthese.
Fokussierte Bibliotheken potentieller Liganden werden
aufgestellt, um kleine organische Moleküle zu suchen, die
spezifisch an ausgewählte Zielproteine binden. So wird die
Analyse der Funktionen und Interaktionen von Proteinen
unterstützt. Natürlicherweise können solche kleinen Moleküle
auch als Leitstrukturen für das Design von Wirkstoffen
dienen. Um die biologische Aktivität von Bibliotheken
kleiner Moleküle untersuchen zu können, hat das FMP
eine Screening Unit etabliert, die Konzepte für das Handling
von Substanzbibliotheken erarbeitet und technische
Voraussetzungen für die Testung Hunderter und Tausender
chemischer Verbindungen schafft.
71 Chemical Biology

PEPTIDE SYNTHESIS
Group Leader: Dr. Michael Beyermann
DEVELOPMENT OF A MODEL FOR LIGAND-
RECEPTOR INTERACTION
Insights into the molecular basis of interaction between
peptide ligands and their membrane-embedded receptors,
in particular G-protein-coupled receptors class B, will
open a way for development of new drugs for treatment
of a number of diseases, since those receptors are very
important targets of peptides regulating many essential
biological functions. Those receptors are functional only
when embedded in the membrane, which is why it is difficult
to obtain direct structural information by spectroscopic
methods such as X-ray crystallography or NMR
methods. Indirect methods, such as structure-activity
relationship studies, modifying the ligand or/and receptor
structure, e.g. by point substitutions, and looking at corresponding
effects on ligand-receptor binding/activation,
may give information about interaction modes between
ligand and receptor. The incorporation of light-directed
crosslinkers into ligands gives the possibility to crosslink
the ligand bound to receptors. By this approach, contact
points between ligand and receptor can be determined.
Furthermore, many evidence points to a crucial contribution
of extracellular receptor domains to ligand binding.
Therefore, we are investigating ligand binding to extracellular
receptor domains. From all these studies, which also
require development of efficient chemical and biotechnological
methods for preparation of receptor domains and
constructs as artificial receptors for appropriate structure
analysis in complex with ligands, we will stepwise
deduce an appropriate interaction model, also taking into
consideration whether receptors interact with ligands as
mono- or oligomers.
CRF receptor antagonists showing different
efficiency (1)
Corticotropin-releasing factor (CRF), urocortin I (Ucn I),
sauvagine (Svg), and urotensin (Uts) are non-selective,
endogenous agonists of the G-protein-coupled receptors
CRF1 and CRF2. Determinants of subtype receptor selectivity,
particularly for such long-chain (~40 aa) peptides, are
widely unknown. First subtype (CRF2) selective agonists
have only been discovered on basis of genomic DNA information
and subsequent cDNA cloning. A strikingly different
amino acid motif (VPIG) at the N-terminus of the
selective agonist Ucn II led to CRF2 selective agonists,
when incorporated into non-selective Ucn I and Svg but
not Utn. Receptor antagonists generated by N-terminal
truncation of CRF2 selective VPIG-Ucn I and Ucn II exhibi-
ted significant CRF2/CRF1 selectivity in terms of affinity
(19 and 260 fold, respectively). Replacing the apparent
selectivity motif VPIG in truncated Ucn II by TFH, residues
at corresponding positions of non-selective Ucn I gave the
most selective antagonist for CRF2, exhibiting an about
1500-fold higher affinity for CRF2 compared to CRF1. Most
remarkably, the CRF2/CRF1 selectivity in terms of antagonistic
potencies and receptor affinity of those antagonists
strongly diverge, although no significant intrinsic activity
was found. The quotient of the relative selectivity in terms
of affinity and potency of a certain antagonist may provide
a basis to evaluate its antagonistic efficiency. The antagonistic
efficiency varied between 2.35 and 33.2 for the
peptides investigated, showing that looking at receptor
affinity only in screenings for antagonists as a global measure
may be insufficient to reflect their inhibitory potency.
Dimerization of corticotropin-releasing factor
receptor type 1 is not coupled to ligand binding
(2)
As reported, receptor dimerization of G-Protein-coupled
receptors may influence signaling, trafficking and regulation
in vivo. Up to now, most studies aiming at the possible
role of receptor dimerization in receptor activation and
signal transduction are focused on class 1 GPCRs. We
have investigated dimerization behavior of CRF1 receptors
tagged with fluorescence labels (CFP/YFP) transiently
expressed in HEK293 cells. We measured Fluorescence
Resonance Energy Transfer and found that CRF1 receptors
form mainly dimers. Upon addition of CRF-related agonists
or antagonists, no significant change of the FRET signal
was observed, indicating no change in the dimer-monomer
ratio by ligand binding.
Photoaffinity cross-linking of the corticotropin-releasing
factor receptor type 1 using photoreactive
urocortin analogues (3)
Interaction of peptide ligands to G-protein-coupled receptors
of class B, such as glucagon, secretin and corticotropin-releasing
factor (CRF), is characterized by a common
orientation of two binding domains, in that ligand C-terminus
binds to extracellular receptor N-terminus (high affinity
binding) and receptor juxta-membrane domain binds
ligand N-terminus. N-terminal truncation, particularly of
residues 6-8 in the case of CRF, leads to antagonists, suggesting
that those residues constitute the receptor activating
sequence. Here, we identified by photoaffinity
cross-linking using Bpa-analogues (Bpa: p-benzoyl-L-phenylalanine)
of urocortin (Ucn) interaction domains of the
receptor (CRF1) with individual amino acids. Specific
digestion patterns of ligand-receptor complexes, obtained
using different cleavage methods and SDS-PAGE for frag-

% Intensity
% Intensity
100
90
80
70
60
50
40
30
20
10
(a)
Spec #1=>BC=>NFO.7=>SM11[BP=666.5, 13430]
941,36
1240,47
1040,41
1339,52
939,39 1141,44
1139,47
1440,53 1640,58
1739,64
1539,60
0
900,0 1162,8 1425,6 1688,4
Mass (m/z)
1951,2 2214,0
100
90
80
70
60
50
40
30
20
10
(b)
ment separation, have shown that 125I-Y-1-Bpa0-Ucn and
125I-Y0-Bpa12-Ucn bind to the second, whilst 125I-Y0-Bpa17- Ucn and 125I-Y0-Bpa22-Ucn bind at the first extracellular
receptor loop. These results show that the suggested activating
sequence, which might penetrate into the transmembrane
segment, is fixed to juxta-membrane domain by
specific interaction of amino acid residues 12, 17 and 22.
Our findings are in conflict with recently published models
in that peptide antagonists bind to receptor N-terminus
enclosing those residues.
Hexa-histidine tag position influences disulfide
pattern (4)
The oxidative folding, particularly the arrangement of disulfide
bonds of recombinant extracellular N-terminal
domains of the corticotropin-releasing factor receptor
type 2a bearing 5 cysteines (C2 to C6) was investigated.
Depending on the position of a His-tag, two types of disulfide
patterns were found. In the case of a N-terminal Histag,
the disulfide bonds C2-C3 and C4-C6 were found, leaving
C5 free, whereas the C-terminal position of the His-tag
led to the disulfide pattern C2-C5 and C4-C6, and leaving
C3 free. The latter pattern is consistent with the disulfide
arrangement of the extracellular N-terminal domain of the
1840,67
Voyager Spec #1=>BC=>NFO.7=>SM35[BP=2042.2, 264]
2042,69
2020,60
1942,18
2003,66
2058,56
1939,71 2040,73
0
800 1300 1800 Mass (m/z) 2300 2800 300
2670,7
264,1
FIGURE 1
MALDI-MS spectra of H-(Val-Thr) 10-NH 2
(crude products) synthesized using standard
Fmoc-chemistry (a) and as the alldepsi
isomer (b) (M[H] + calc: 2019.18)
CRF receptor type 1, which has six cysteines (C1 to C6) and
in which C1 is paired with C3. However, binding data of the
two differently disulfide-bridged domains show no significant
differences in binding affinities to selected ligands,
indicating the importance of the C-terminal portion of the
N-terminal receptor domains, particularly the disulfide
bond C4-C6 for ligand binding.
An unexpected side reaction gets a beneficial
tool: N↔O-acyl shifts (5)
N-to-O acyl migrations at serine or threonine residues in
peptides or proteins have previously been observed upon
treatment with strong acids. We observed such shifts, surprisingly,
for the moderately strong acid trifluoroacetic
acid (TFA) used in Fmoc chemistry for the final removal of
all protecting groups. N,O-acyl shifts have not been recognized
as leading to serious side reactions in Fmoc-based
peptide synthesis and, thus, are not even mentioned in
recent textbooks. The extent of undesired N→O shifts
depends on the peptide sequence and even in the case of
TFA may give rise to large amounts of depsipeptide by-products.
Moreover, we found that syntheses of sequences
in which the ester unit appears at appropriate positions
were far more efficient than the syntheses of those with-
Chemical Biology
73

out the ester unit. A final ammonium hydroxide-induced
O→N-shift yielded the authentic peptides in much better
yields. This new method opens a way for efficient synthesis
of difficult sequences, thus facilitating investigations
on the role of individual amino acid residues on structural
stability of the WW domain FBP-28. Compared with other
methods, such as pseudoproline methodology, an important
advantage of the depsipeptide method results from
the fact that the conformation-disrupting modification
introduced by the depsipeptide unit is still present during
chromatographic purification of the deblocked peptide.
The suppression of conformation-induced association
leads to an increased solubility and narrower peaks for
depsi isomers, thus allowing a much more efficient purification.
Group members
Dr. Jana Klose (4)
DC Angelika Ehrlich (SPOT library synthesis)*
Doreen Wietfeld (Doctoral student) (1)*
Sandra Tremmel (Doctoral student) (5)
Oliver Krätke (Doctoral student) (2,3)
Stephan Pritz (Doctoral student)*
Irene Coin (Doctoral student) (5)*
Annerose Klose (Technical assistance peptide chemistry)
Dagmar Krause (Technical assistance peptide purification,
analysis)
Barbara Pisarz (Technical assistance Biacore)**
Bernhard Schmikale (Technical assistance peptide synthesis)**
External funding
Deutsche Forschungsgemeinschaft
Projekt im Schwerpunktprogramm „Molekulare Sinnesphysiologie“
(SPP 1025)
Michael Beyermann
Deutsche Forschungsgemeinschaft
„Untersuchungen von CRF- und CRF-Rezeptor-Mutanten
zur Entwicklung eines Modells für die Ligandenerkennung
von G-Protein-gekoppelten Rezeptoren der Familie 2“ (TP
A6 im Sonderforschungsbereich 449 „Struktur und Funktion
membranständiger Rezeptoren“)
Michael Bienert, Michael Beyermann, Walter Rosenthal
Deutsche Forschungsgemeinschaft
„Organisation und Funktion eines Transduktionskomplexes
in Sehstäbchen“ (BE 1434/3-3)
Michael Beyermann
* part of period reported
** part-time
Deutsche Forschungsgemeinschaft
„Struktur, Stabilität und Spezifikationen von nichtkatalytischen
Proteindomänen und deren Verwendung als
Werkzeuge für das Design einer stabilen minimalen ß-Faltblattstruktur
und das Verständnis von pathologischen Prozessen“
(TP 2/2-1 in der Forschergruppe 299 „Optimierte
molekulare Bibliotheken zum Studium biologischer Erkennungsprozesse“)
Hartmut Oschkinat, Michael Bienert
Selected publications (FMP authors in bold)
Klose J, Fechner K, Beyermann M, Krause E, Wendt N,
Bienert M, Rudolph R, Rothemund S (2005) Impact of
N-terminal domains for corticotropin-releasing factor
(CRF) receptor-ligand interactions. Biochem, in press
Klose J, Wendt N, Kubald S, Krause E, Fechner K, Beyermann
M, Bienert M, Rudolph R, Rothemund S (2004) Hexahistidin
tag position influences disulfide structure but not
binding behavior of in vitro folded N-terminal domain of rat
corticotropin-releasing factor receptor type 2a. Protein Sci
13, 2470-2475
Carpino LA, Krause E, Sferdean CD, Schümann M, Fabian
H, Bienert M, Beyermann M (2004) Synthesis of ”Difficult”
Peptide Sequences: Application of a Depsipeptide Technique
to the Jung-Redemann 10- and 26-mers and the
Amyloid Peptide A‚(1-42). Tetrahedron Lett 45, 7519-7523
Eggelkraut-Gottanka R, Klose A, Beck-Sickinger AG, Beyermann
M (2003) Peptide (alpha)thioester formation using
standard Fmoc-chemistry. Tetrahedron Lett 44, 3551-3554
Kaupp UB, Solzin J, Hildebrand E, Brown JE, Helbig A,
Hagen V, Beyermann M, Pampaloni F, Weyand I (2003) The
signal flow and motor response controlling chemotaxis of
sea urchin sperm. Nature Cell Biol 5, 109-117
Wietfeld D, Fechner K, Heinrich N, Seifert R, Bienert M,
Beyermann M (2003) Development of CRF receptor-2
selective peptide ligands. Biopolymers 71, 376
Collaborations
In a number of projects we have collaborated either with
groups within and/or outside the FMP, particularly with
L.A. Carpino (University of Massachusetts/USA) on development
of a new method for synthesis of difficult peptides,
with A.G. Beck-Sickinger (University of Leipzig) on preparation
and use of peptide thiol esters for peptide and protein
synthesis, and with U.B. Kaupp (FZ Jülich)/V. Hagen
(FMP) on studies of chemotaxis using appropriately caged
peptides. Of particular impact is the joint work with the
Peptide Biochemistry Group.

PEPTIDE LIPID INTERACTION/PEPTIDE
TRANSPORT
Group Leaders: Dr. Margitta Dathe and PD Dr. Johannes
Oehlke
PROPERTIES OF CELLULAR
UPTAKE-MEDIATING PEPTIDES
The application of many potential, biotechnologically available
drugs of e.g. proteinaceous or nucleic acid origin is
limited by their inefficient cellular uptake caused by the
low permeability of the cell membrane. Commonly used
uptake-promoting carrier systems based on cationic lipids
or virus components are hampered by low efficiency, poor
specificity, poor bioavailability, toxicity and immunological
risks. Thus, promising new drugs will not be introduced
in diagnostics and therapy if we are not able to develop
safe, efficient application forms. As an alternative,
during the last decade small peptide carriers designated
as cell-penetrating peptides (CPP) or protein transduction
domains (PTDs) have been introduced as uptake-facilitating
compounds. Covalent linkage or complexation of
such peptides with bioactive polymers up to the size of
proteins and nucleic acids led to highly efficient delivery
into and across a variety of cells, thereby avoiding the
drawbacks of the common delivery systems. Peptide
coupling to potential drug containers such as liposomes
which can be loaded with hydrophobic as well as hydrophilic
compounds provide the additional advantage of
transport of large amounts of drug molecules with unfavorable
properties such as poor solubility and low stability.
Since many of the peptides promote membrane-reorientation
processes involving temporary membrane destabilization
and energy-independent uptake, it has been suggested
that the lipid bilayer of cell membranes is targeted.
Peptides which contain recognition motifs suitable for
addressing well-defined receptor structures at the cell
membrane provide an opportunity for site-specific delivery.
The main goal of our group is the elucidation of the structural
requirements for peptides to deliver covalently or
complex-bound drugs across cell membranes by nonendocytotic
as well as endocytotic mechanisms. The
research efforts in this context are focused on studying
interactions of CPPs with lipid membranes to get mechanistic
insights into transmembrane transport routes and on
the development of peptide-liposome complexes for drug
delivery to the brain (M. Dathe). As a further topic, the
structural requirements for the delivery ability of CPPs are
being studied, using covalently bound peptide nucleic
acids (PNAs) as cargo molecules by pursuing the uptake
into different cell types in comparison with the biological
effects of the PNAs in various systems (J. Oehlke). Additionally,
we are interested in elucidating the structural principles
and the mechanism of action of peptides which
selectively kill bacteria by destroying their cell membrane.
These studies aim at contributing to the development of a
new class of peptidic antibiotics (M. Dathe).
Cellular uptake of PNA after conjugation with
positively and negatively charged ∝-helical
amphipathic, β-sheet forming and unstructured
peptides
Yvonne Wolf, Angelika Ehrlich, Burkhard Wiesner, Michael
Bienert, and Johannes Oehlke
In order to contribute to an elucidation of the structural
requirements for the shuttling ability of cell-penetrating
peptides, we investigated the cellular uptake of a 12-mer
PNA directed against the mRNA of the nociceptin/orphanin
FQ receptor after disulfide bridging with various peptides
showing different structure forming properties, charge
and size (Table). As the lead we used an amide-bound
conjugate of the PNA with the cell-penetrating α-helical
amphipathic model peptide KLALK LALKA LKAAL KLA-NH 2
(KLA). We have shown previously that KLA exhibits up to
tenfold higher cellular uptake and biological activity than
the naked PNA. The cellular uptake was studied by means
of capillary electrophoresis combined with laser-induced
fluorescence detection (CE-LIF). Using this approach a
separate quantification of the conjugate and the naked
PNA generated by cleavage of the disulfide bond in the
reducing environment of the cell interior was possible,
thus avoiding bias of the results by surface adsorption.
Assessment of the cellular uptake into CHO cells and
cardiomyocytes by CE-LIF revealed enrichments within the
cell interior of about two and tenfold for the naked PNA
and its amide bound KLA conjugate, respectively (related
to the external concentration and an experimentally determined
ratio of about 10 µl cell volume/mg protein). Confocal
laser scanning microscopy (CLSM) and fluorescence
activated cell sorting (FACS) supported these results. An
extensive cellular uptake was found also for the analogous
disulfide bridged KLA-PNA-conjugate. But, for reasons
which still remain unclear, the intracellular PNA concentration
in this case only approached that provided externally.
Even strong structural alterations in the peptide part
of the disulfide bridged conjugate did not seriously influence
the extent of uptake into HEK- and CHO cells (Fig. 1),
contradicting speculations about specific structural requirements
for the shuttling ability of peptides. Surprisingly,
after energy depletion an enhanced uptake into CHO cells
as well as into HEK cells was found for conjugates bearing
Chemical Biology
75

Compound Sequence Structural properties
Peptide nucleic acid Fluos-GGA GCA GGA AAG-Cys
KLA Dansyl-G-C-KLALK LALKA LKAAL KLA-NH2 α-helical, amphipathic
KGL Dansyl-G-C-KGLKL KGGLG LLGKL KLG- NH 2 unstructured
ELA Dansyl-G-C-ELALE LALEA LEAAL ELA-NH 2 α-helical, amphipathic
RLA Dansyl-G-C-RLALR LALRA LRAAL RLA-NH 2 α-helical, amphipathic
Penetratin Dansyl-G-C-RQIKI WFQNR RMKWK K-NH 2 poor α-helical amphipathic
VT5 Dansyl-G-C-DPKGDPKGVTVTVTVTVTGKGDPKPG- NH 2 β-sheet, amphipathic
TABLE 1
Components of disulfide-bridged PNA-peptide conjugates.
an acidic helical amphipathic and a ß-sheet forming peptide,
respectively (Fig. 1). It can be inferred from the latter
findings that the uptake under normal conditions can be
counteracted differently by active transport systems in
dependence on both the peptide and the cell type, thus
providing a potential basis for achieving cell selective
cargo delivery.
Peptide transport across lipid bilayers
S. Keller, E. Bárány-Wallje 1, S. Serovy 2, M. Bienert, M.
Dathe
Conflicting reports on the translocation ability of the cellpenetrating
peptides through pure lipid bilayers have
stimulated biophysical studies on the interaction of penetratin,
a peptide capable of transporting large hydrophilic
cargos into the cell with model lipid membranes to compare
its bilayer behavior with the ability to enter cells. The
results, which provided no evidence for peptide translocation
through pure lipid bilayers, support suggestions
that the highly cationic peptide enters living cells by an
endocytotic pathway rather then by disturbing the integ-
1) Dept. of Biochemistry and Biophysics, Stockholm University
2) Junior Research Group Biophysics
rity of the lipid matrix and formation of transient and local
lipid structures.
Brain targeting by apolipoprotein E-peptideliposome
complexes
I. Sauer, O. Liesenfeld 3, H. Scharnagel 4 , K. Weisgraber 5,
M. Bienert, M. Dathe
The treatment of central nervous system diseases is a particular
challenge. Different from blood vessels of other
organs which are equipped with small pores, the tightly
connected brain capillary endothelial cells forming the
blood-brain barrier (BBB) prevent paracellular transport.
Furthermore, most potential drugs and modern molecular
tools are neither able to diffuse across the cell layer nor
are accessible to the specialized receptor-mediated transport
systems. An approach to overcome unfavorable properties
provides for drug incorporation into carriers. Liposomes
have emerged as a very potential drug container.
3) Institute for Infectious Medicine, Charité Campus Benjamin Franklin,
Berlin
4) Dept. of Clinical Chemistry, University Hospital Freiburg im Breisgau
5) Gladstone Institute of Cardiovascular Disease, San Francisco
6) NMR-Spectroscopy Group, FMP

FIGURE 1
Quantities of PNA-Cys in the extracts of HEK- and CHO-cells exposed before to 0.2 µM PNA alone or disulfide bridged
with various peptides (table) for 60 min at 37 °C without (normal) and with energy depletion (after 60 min preincubation
with 2 deoxy-glucose/Na-azide 25/10 mM). Each bar represents the mean of three samples + SEM.
The solvent-containing interior of the nanoscopic particles
self-assembled from lipids can be loaded with polar drugs
and the hydrophobic shell with non-polar compounds.
Equipped with an appropriate transport-mediating compound
that recognizes membrane constituents of brain
capillary endothelial cells, thousands of incorporated drug
molecules could be transported.
It was our objective (M. Dathe and coworkers) to develop
peptidic vectors which recognize the low density lipoprotein
receptor (LDL receptor) on brain capillary endothelial
cells, and to develop strategies of their efficient coupling
to liposomal carriers. The receptor was found to be highly
expressed on capillary endothelial cells of different species.
A peptide encompassing the highly cationic tandem
dimer peptide (141-150)2 derived from the LDL receptorbinding
domain of apolipoprotein E (apoE) was used as
vector sequence. A cost-effective approach to coat liposomes
with an apoE peptide and to induce the structure
that enables binding to the LDL receptor relies on the
adsorption to the liposomal surface. Amphipathic and
transmembrane helices and fatty acid chains served as
anchoring domains. Since coating vesicle surfaces with
peptides could promote the destabilization of liposomal
preparations rendering vesicles leaky and peptides
released in circulation could cause perturbation of cell
membranes, their stable anchorage at the liposomal surface,
the potential to assume a receptor-recognizing helical
conformation, their ability to conserve the integrity of
the vesicles, and their toxic effect were evaluated.
Complexes of liposomes with a dipalmitoyl-modified apoE
peptide proved to be the most promising to combine the
purposes of a stable drug container and targeting to brain
capillary endothelial cells. Furthermore, covalent coupling
of the cationic apoE peptide onto sterically stabilized liposomes
provide stable complexes.
The peptide-coated liposomes were efficiently internalized
into endothelial cells of brain microvessels by an energydependent
endocytotic process (Figure 1). Low peptide
affinity to the LDL receptor and internalization into cells
with up- and down-regulated receptor level pointed to the
dominating role of an LDL receptor-independent transport
route. Studies to influence unspecific charge interaction
of the cationized liposomes with anionic membrane constituents
suggested that the ubiquitously expressed cell
matrix component heparan sulfate proteoglycan (HSPG)
played a decisive role in the uptake process.
Chemical Biology
77

The efficient uptake via HSPG-involving endocytosis may
provide a promising strategy for facilitated drug delivery
across the blood brain barrier. Furthermore, the observed
internalization emphasized the potential of cationic peptides
to ferry complex drug carriers into cells, and the
adsorptive coupling strategy offers great adaptability to a
broad spectrum of ligands destined to diverse biological
targets.
Group members
Ines Sauer (Doctoral student)
Sandro Keller (Doctoral student)*
Yvonne Wolf (Doctoral student)*
Axel Wessolowki (Doctoral student)*
Heike Nikolenko (Technical assistance)
Gabriela Vogelreiter (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
“Linear and cyclic hexapeptides: interaction with membranes
and modulation of selective cell-lytic processes” (DA
324/4-1, 4-2)
Margitta Dathe
* part of period reported
FIGURE 2
Confocal laser scanning microscopic picture of mouse
brain capillary endothelial cells exposed to fluorescence-labeled
and apoE peptide-covered liposomes at
T = 37 °C for t = 30 min. Uptake is documented by the
spotted fluorescence within the cytoplasm.
Deutsche Forschungsgemeinschaft
“Passage of the blood-brain barrier using surface-modified
nanosuspensions and apolipoprotein-E peptidecovered
carrier systems” (DA 324/5-1)
Margitta Dathe
Deutsche Forschungsgemeinschaft
“Hirntargeting mittels oberflächenmodifizierter Nanosuspensionen
und Apolipoprotein E-Peptid beladener
Trägersysteme“ (Teilprojekt 7B der Forschergruppe 463
“Innovative Arzneistoffe und Trägersysteme – Integrative
Optimierung zur Behandlung entzündlicher und hyperproliferativer
Erkrankungen“)
Margitta Dathe
European Community
“Investigation of structural prerequisites for cell-selective
drug uptake mediated by peptide vectors” (Subproject of
QLK3-CT-2002-01989 “Target specific delivery systems for
gene therapy based on cell penetrating peptides”)
Johannes Oehlke, M. Bienert
European Community
“Interaction of cell-penetrating with artificial lipid membranes
and peptide translocation through lipid layers”
(Subproject of QLK3-CT-2002-01989 “Target specific deli-

very systems for gene therapy based on cell penetrating
peptides”)
Margitta Dathe, Michael Bienert
Deutscher Akademischer Austauschdienst
“Untersuchungen zu strukturellen Voraussetzungen für
einen zellselektiven Wirkstofftransport durch Peptidvektoren“
(DAAD-Stipendium – 3-monatiger Studienaufenthalt an
der Universität Montpellier, Frankreich)
Yvonne Wolf
Deutscher Akademischer Austauschdienst
“Interaction of amphipathic ApoE-peptides with membranes:
modulation of a receptor-mediated drug transport into
the brain”
(DAAD-Stipendium: 4-monatiger Studienaufenthalt am
J. David Gladstone Institute, San Francisco, USA
Ines Sauer
Deutscher Akademischer Austauschdienst
“Lineare und cyclische Hexapeptide: Interaktion mit Membranen
und Modulation selektiver zelllytischer Prozesse“
(DAAD-Stipendium: 2-monatiger Studienaufenthalt am
Karolinska Institut, Stockholm, Schweden)
Axel Wessolowski
Organizing committee of the Congress “Cellular Transport
Strategies for Targeting of Epitopes, Drugs and Reporter
Molecules“, (Budapest 2003), Conference grant
Ines Sauer
Selected publications (FMP authors in bold)
Dathe M, Nikolenko H, Klose J, Bienert M (2004) Cyclization
increases the antimicrobial activity and selectivity of
arginine- and tryptophan-containing hexapeptides. Biochemistry
43, 9140-9150
Wessolowski A, Bienert M, Dathe M (2004) Antimicrobial
activity of arginine- and tryptophan-rich hexapeptides: the
effect of aromatic clusters, D-amino acid substitution and
cyclization. J Pept Res 64, 159-169
Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B, Berger
H, Bienert M (2004)
Enhancement of intracellular concentration and biological
activity of PNA after conjugation with a cell-penetrating
synthetic model peptide. Eur J Biochem 271, 3043-3049
Hällbrink M, Oehlke J, Papsdorf G, Bienert M (2004)
Uptake of cell-penetrating peptides is dependent on the
peptide-to-cell-ratio rather than on peptide concentration.
Biochim Biophys Acta 1667, 222-228
Sauer I, Dunay IR, Weisgraber K, Bienert M, Dathe M
(2005) An apolipoprotein E-derived peptide mediates
uptake of sterically stabilized liposomes into brain capillary
endothelial cells. Biochemistry 44, 2021-2029
Collaborations
Prof. Oliver Liesenfeld, Dept. of Medical Microbiology and
Immunology of Infection, Charité Campus Benhamin
Franklin, Berlin
Dr. Hubert Scharnagel, Dept. of Clinical Chemistry, University
Hospital Freiburg im Breisgau
Prof. Karl Weisgraber, Gladstone Institute of Cardiovascular
Disease, San Francisco
Chemical Biology
79

PEPTIDE BIOCHEMISTRY
Group Leader: Dr. Hartmut Berger
CORTICOTROPIN-RELEASING FACTOR
RECEPTOR TYPE 1 (CRFR1): REGULATION
OF THE COUPLING TO DIFFERENT G-PRO-
TEINS AND THE TWO-DOMAIN RECEPTOR
MODEL
Corticotropin-releasing factor (CRF) receptor type 1
(CRFR1) is a G-protein-coupled receptor (GPCR) of the
class B, secretin receptor family, which is activated by
several peptide ligands and which is thought to be the
principal physiological mediator of stress responses. For
the activation of the receptor by peptide ligands, a twodomain
model has been established by several authors:
the N-terminal domain of the receptor (N-domain) is required
for ligand binding, whereas the juxtamembrane
region, consisting of transmembrane regions and intervening
loops (J-domain), is involved in the activation of the
signaling steps by the ligand, bitethered between N and J.
Furthermore, whereas peptide antagonists bind predominantly
to the N-domain with high affinities, non-peptide
antagonists bind only to the J-domain. We have provided
evidence that the CRFR1 expressed in HEK (HEK-CRFR1)
cells couples to G s- and G i-proteins with different characteristics.
The two-domain binding model and our model on
G-protein coupling regard different aspects of the activation
of the CRFR1. Therefore, it is important to investigate
whether they can be fitted into one combined model.
For this reason, we studied the regulation of G-protein
coupling of the CRFR1 in HEK-CRFR1 cell membranes and
the influence of peptide and non-peptide antagonists on
the coupling of the receptor, using the GTPγ 35S binding
assay (Hartmut Berger, Nadja Heinrich, Monika Georgi,
Doreen Wietfeld*, Jens Furkert**). Corresponding to
biphasic binding of the ligand to the CRFR1, G s- and G i-protein
coupling of the receptor were seen in the biphasic
ligand-evoked stimulation of GTPγ 35S binding to HEK-
CRFR1 membranes (Fig. 1). G s-activation could be separated
by inactivation of G i with pertussis toxin, G i activation
by pre-stimulation of the cells with a ligand, which only
desensitized Gs activation (Fig. 1). The specificity of G s and
G i activation was confirmed by immunoprecipitation of
GTPγ 35S-bound G α subunits. By this means and a substantial
accumulation of inositol phosphates (IP3) in cells, it
was found that the receptor also activates G q-proteins.
* Peptide Synthesis Group,
**Cellular Biology/Molecular Medicine Group
Studying the characteristics of the coupling of CRFR1 to
G s, G i, and G q in more detail, we developed the following
model for the activation of the signaling by CRFR1 in HEK
cells. A peptide agonist ligand binds to a high-affinity, lowcapacity
receptor binding site activating G s-protein and
adenylate cyclase with high ligand potency. At higher concentration
the ligand additionally binds to one or more lowaffinity,
high-capacity sites of which the main part remains
non-coupled, however a small number couples to G i-proteins,
attenuating the ligand-stimulated production of
cAMP. The activation of the G-proteins is accomplished by
increasing the affinity of the nucleotide-binding site in the
G α chain for GTP, the affinity of GTPγS to G i being stronger
increased as compared to G s. Very high ligand concentrations
also stimulate IP3 accumulation through activation
of G q and, in addition, through a pertussis toxin-dependent
way, obviously by activation of phospholipase C by the G βγ
chain released from G i when G i is activated. Stimulation of
the receptor by ligands desensitizes G s and G q activation,
but not G i, so that the inositol phosphate pathway is about
half-desensitized.
From our findings it seems clear that the CRFR1 can be
added to the growing list of GPCRs that simultaneously
couple to unrelated G-proteins, possibly through different
active receptor states and/or different affinities of the
G-proteins to one or more such states. To get more insight
into these possibilities, we studied the influence of peptide
and non-peptide antagonist on the G-protein coupling,
on the basis of the two-domain model for CRFR1. The peptide
antagonist α-helical CRF(9-41), assumed to bind predominantly
to the N-domain, antagonized G s as well as G i
coupling competitively with equal potencies, suggesting
that similar ligand binding to N is responsible for G s as well
as G i activation. However, the non-peptide antalarmin,
assumed to bind exclusively to the J-domain, antagonized
Gs activation competitively but the G i response non-competitively
(Fig. 2). This should mean that nonpeptide antagonists
antagonize G s and G i coupling of CRFR1 by competitive
and allosteric mechanisms, respectively, and that
different conformation ensembles of two possibly overlapping
J domains of the receptor are responsible for the
coupling to G s and G i.
Taken together, it is concluded that CRFR1 signals through
G s-, G i-, and G q-proteins. The concentrations of the stimulating
ligand and GTP and, furthermore, selective desensitization
may be part of a regulatory mechanism determining
the actual ratio of the coupling of CRFR1 to different
G-proteins. G s and G i coupling of the receptor can be described
within the framework of the two-domain receptor

FIGURE 1
G s- and G i-protein coupling of CRF
receptor type 1 as measured by sauvagine-stimulated
binding of GTPγ 35S to
membranes obtained from HEK cells
expressing the receptor.
FIGURE 2
Competitive and noncompetitive antagonism
by the non-peptide antalarmin of,
respectively, G s and G i coupling of the
CRF receptor type 1 expressed in HEK
cells. G-protein coupling was measured
as sauvagine-stimulated binding of
GTPγ 35S to membranes obtained from
the cells after inactivation of G i by pertussis
toxin (Gs activity left, A) or desensitization
of Gs coupling by sauvagine
(G i activity left, B).
Chemical Biology
81

model in that they are accomplished by different active
conformations of the J-domain.
Our group was also engaged in receptor assays for studying
structure-activity relationships of CRF peptides (Klaus
Fechner, Gabriela Vogelreiter, results are given by the Peptide
Synthesis Group).
Group members
Dr. Nadja Heinrich**
Dr. Klaus Fechner*
Monika Georgi (Technical assistance)
Gabriela Vogelreiter (Technical assistance)**
Selected Publications (FMP authors in bold)
Wietfeld D, Heinrich N, Furkert J, Fechner K, Beyermann
M, Bienert M, Berger H (2004) Regulation of the coupling
to different G proteins of rat corticotropin-releasing factor
receptor type 1 in human embryonic kidney 293 cells. J
Biol Chem 279, 38386-38394
Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B, Berger
H, Bienert M (2004) Enhancement of intracellular concentration
and biological activity of PNA after conjugation
with a cell-penetrating synthetic model peptide. Eur J Biochem
271, 3043-3049
* part of period reported
** part-time

MASS SPECTROMETRY
Group Leader: Dr. Eberhard Krause
MASS SPECTROMETRY-BASED PROTEIN
ANALYSIS
Recent results emphasize the role of mass spectrometry
as an essential tool for proteom studies in the field of cellular
biology. The identification and quantitation of very
complex protein mixtures as well as of low abundance
signaling proteins require highly sensitive analytical techniques.
Based on the methodological advances in peptide
and protein ionization (awarded with the Nobel Prize in
2002) and new powerful fragmentation techniques
(MS/MS), state of the art mass spectrometry is able to provide
very accurate and sensitive mass measurements as
well as sequence information of peptides and high molecular
weight proteins. The MS group contributed to several
proteom projects which include the identification of
specific proteins of molecular networks in cells following
functional stimulation and the elucidation of functionally
important post-translational modifications of proteins. In
order to gain insight into the ionization behavior of peptides
and proteins, the group is performing methodological studies.
Using chemical derivatization strategies and stable
isotope-labeling, improved methods are being developed
for the MS-based analysis of phosphoproteins and for the
high sensitive identification and quantitation of low-expressed
signaling proteins, respectively. Both the proteomics
work and the methodological studies are being carried out
in collaboration with groups from both within and outside
the FMP.
Examination of erythropoietin receptor functions
using different proteomic strategies
The interaction between erythropoietin (EPO) and its
receptor, which is expressed on late erythroid progenitor
cells, initiates multiple signaling pathways by the recruitment
of cytosolic src homology 2 (SH2) domain-containing
proteins to the phosphorylated receptor. The phosphorylation
of receptor tyrosine residues is mainly due to the
activity of the Janus family kinase 2 (JAK2), a receptorassociated
enzyme which is undergoing transphosphorylation
after receptor dimerization. Studies using mutated
forms of the EPO receptor (EPOR) have identified single
tyrosine residues that are critical for the recruitment of
distinct signaling proteins. However, it is still difficult to
relate the function of single tyrosine residues/receptor
subdomains or distinct signaling molecules/pathways to
the induction of proliferative, antiapoptotic or differentiative
cellular responses initiated after the binding of EPO to
its receptor. Proteomic strategies seem to have an enor-
mous potential in the analysis of changes in the abundance
of proteins, but also in their post-translational modification.
A major challenge in creating phosphoproteome
maps, however, remains the fact that phosphorylated
regulatory molecules are expressed at rather low levels.
We applied different proteomic strategies to the investigation
of structure-function relationships in the EPOR signaling
complex. Using a set of EPOR mutants expressed in an
identical cellular background (Fig. 1), we focused on rapid
changes in the tyrosine phosphorylation state of proteins.
However, the analysis using the classical approach combining
two-dimensional gel electrophoresis (2-DE) and
identification by MALDI-MS revealed that low expressed
signaling proteins cannot be detected by this technique. An
alternative strategy using 1-D gel separation of phosphoproteins
and LC-tandem mass spectrometry (MS/MS)
allowed us to identify proteins which are involved in intracellular
signaling. Besides proteins of already established
pathways, proteins could be identified which have not been
linked to EPO-induced signalling so far. Thus, the phosphorylation
of SUMO-1 has been suggested to be part of strategies
repressing cytokine induced signals in analogy to
the covalent ubiquitin attachment. We identified several
GTP binding proteins as well as proteins involved in the
modification/regulation of G-proteins which have not been
reported to be involved in EPOR induced regulatory events.
In addition, a stable isotope labeling method (Fig. 2) was
used for quantitative analysis of gel-separated proteins of
the EPOR signaling. The results show that identification
and relative quantification of more than 200 EPOR-dependent
proteins could be achieved. We can conclude from
our experiments that a proteomic strategy based on a
combination of one-dimensional gel separation, in-gel
18O-labeling, and LC-MS/MS provides the sensitivity
required for the detection of low expressed signaling
molecules and offers the potential for a more comprehensive
analysis of complex cellular responses (cooperation
with T. Bittorf and S. Körbel, Institute for Medical Biochemistry
and Molecular Biology, University of Rostock).
Chemical derivatization strategies for analysis
of post-translational modifications of proteins
Esterification of the hydroxyl functions of threonine, serine,
and tyrosine with a phosphate group is the ubiquitous
modification of proteins which is involved in many signal
transduction processes, for example, cell differentiation,
proliferation, energy storing, and apoptosis. Although
mass spectrometry has become a preferred method to
analyze phosphorylated proteins, the determination of the
site of phosphorylation, especially for high molecular
weight proteins, remains far from routine. The low abun-
Chemical Biology
83

FIGURE 1
Schematic representation of wild-type
and mutated EGF/EPO receptors. All
receptors consist of the human EGFR
ligand-binding domain and the transmembrane
and cytoplasmatic domains
of the murine EPOR.
FIGURE 2
Scheme for the identification and relative
quantitation of EPOR-dependent
phosphoproteins by enzymatic isotope
labeling.

dance of phosphoproteins, the difficulty of obtaining full
sequence coverage by specific proteolysis, and the low
ionization efficiency of phosphopeptides compared with
their non-phosphorylated analogs may prevent the detection
of specific phosphopeptides. In our group, the betaelimination/Michael
addition reaction was used to replace
the phosphate moiety of phosphoserine or phosphothreonine
peptides by a group which should give rise to remarkably
enhanced signal intensities in MALDI mass spectrometry.
Several N- and S-nucleophiles were studied with
regard to enhanced ionization efficiency, and optimized
reaction conditions for beta-elimination/addition procedures
were developed. Based on in-gel derivatization with
2 phenylethanethiol (PET) we demonstrate that enhanced
ionization efficiencies can be used for sensitive MS analysis
of protein phosphorylation. This method was applied
to determine the phosphorylation of Stat1. Stat proteins
(signal transducers and activators of transcription) are
cytokine-responsive transcription factors which play an
important role in the regulation of gene expression. Several
post-translational modifications are required for activation,
whereas serine phosphorylation at position 727 is
necessary for transcriptional activity. The derivatization
allowed direct MS detection of the peptide sequence
covering the phosphoserine position 727 of Stat1, which is
normally suppressed in complex peptide mixtures.
In addition, mass spectrometric analysis was able to provide
evidence contradicting recent results on the role of
Arg31 methylation in the DNA binding of Stat1. MS and
tandem MS measurements clearly revealed that arginine
in position 31 of Stat1 is not methylated to a significant
extent. In conclusion, alternative explanations to methylation
have to be explored to understand the molecular
mechanism of reduced interferon sensitivity of tumor cells
which accumulate high levels of the methyltransferase
inhibitor methylthioadenosine (in cooperation with T.
Meissner and U. Vinkemeier, FMP).
Group members
Dr. Michael Schümann
Clementine Klemm (Doctoral student)
Heidi Lerch (Technical assistance)
Kareen Tenz (Technical assistance)*
Stephanie Lamer (Technical assistance)*
External funding
Deutsche Forschungsgemeinschaft
„Simultane Untersuchung Erythropoietinrezeptor-abhängiger
Signale durch funktionelle Proteomanalyse“ (BI 599/2)
Thomas Bittorf (University of Rostock), E. Krause
* part of period reported
Bundesministerium für Bildung und Forschung
„Identifizierung hepatozellulärer Biomarker“ (0312618/UA
Massenspektrometrie) Eberhard Krause
Selected publications (FMP authors in bold)
Kleuss C, Krause E (2003) G alpha s is palmitoylated at the
N-terminal glycine residue. EMBO J 22, 826-832
Czupalla C, Culo M, Müller EC, Brock C, Reusch HP,
Spicher K, Krause E, Nürnberg B (2003) Identification and
characterization of the autophosphorylation sites of phosphoinositide
3-kinase isoforms beta and gamma. J Biol
Chem 278, 11536-11545
Kraus M, Bienert M, Krause E (2003) Hydrogen exchange
studies on Alzheimer’s amyloid-beta peptides by mass
spectrometry using matrix-assisted laser desorption/ionization
and electrospray ionization. Rapid Commun Mass
Spectrom 17, 222-228
Carpino LA, Krause E, Sferdean CD, Schümann M, Fabian
H, Bienert M, Beyermann M (2004) Synthesis of “difficult“
peptide sequences: application of a depsipeptide technique
to the Jung-Redemann 10- and 26-mers and the amyloid
peptide Abeta(1-42). Tetrahedron Lett 45, 7519-7523
Baumgart S, Lindner Y, Kühne R, Oberemm A, Wenschuh
H, Krause E (2004) The contributions of specific amino acid
side chains to signal intensities of peptides in matrix-assisted
laser desorption/ionization mass spectrometry. Rapid
Commun Mass Spectrom 18, 863-868
Klemm C, Schröder S, Glückmann M, Beyermann M,
Krause E (2004) Derivatization of phosphorylated peptides
with S- and N-nucleophiles for enhanced ionization efficiency
in matrix-assisted laser desorption/ionization mass spectrometry.
Rapid Commun Mass Spectrom 18, 2697-2705
Meissner T, Krause E, Lödige I, Vinkemeier U (2004) Arginine
methylation of STAT1: a reassessment. Cell 119, 587-589
Collaborations
Thomas Bittorf, University of Rostock
Phosphoproteomics of EPO receptor signaling
Ursula Gundert-Remy, Axel Oberemm, BfR Berlin
Proteom analysis for risk assessment of chemicals
Christiane Kleuss, Institute for Pharmacology, Charité –
University Medicine Berlin
Modifications of G-proteins
Louis A. Carpino, University of Massachusetts, Amherst,
MA, USA
Side reaction in peptide synthesis
Chemical Biology
85

SYNTHETIC ORGANIC BIOCHEMISTRY
Group Leader: Dr. Volker Hagen
CAGED COMPOUNDS
Controlled temporal and spatial release of biomolecules
from photolabile precursors, called caged compounds,
have become increasingly interesting in the chemical and
biological communities. When caged, a biomolecule is
rendered biologically inactive by derivatization with a
photolabile protecting or caging group. As a result, it can
be applied to cells under steady state conditions without
evoking biological responses. Flash photolysis using UV
light cleaves the modifying group and rapidly generates
the biologically active molecule. Because photolysis of
caged compounds generates effectors in situ, much faster
and more spatially uniform, concentration jumps can be
triggered than with other techniques. There are no diffusion
problems and no spatial inhomogeneity problems
with addition of substrates. Desensitization is minimized.
Caging and uncaging of biomolecules are widely used for
studies of mechanisms and the kinetics of cellular processes.
Other applications of photoremovable protecting
groups which are currently being explored are time resolved
X-ray studies, protein folding, gene expression control,
and also combinatorial chemistry and photolithography.
Caged compounds must meet specific requirements. They
should undergo fast and highly efficient photochemical
reactions and display high molar absorptivities at wavelengths
>300 nm, or better >350 nm. Furthermore, they
should be sufficiently soluble in aqueous solutions, stable
towards solvolysis, and biologically inert. Sometimes
membrane-permeability is required. Commonly used
caged compounds do not meet these requirements, therefore
the design of novel caging groups and the development
of novel caged biomolecules are necessary. Furthermore,
a comparatively small number of caged compounds
exists at present and the variety of applications is such
that it is desirable to have a range of compounds with different
characteristics to choose from.
Our group deals with the development of novel caging
groups and the synthesis, characterization and application
of caged compounds. In the past few years, we developed
[5,7-bis(carboxymethoxy)coumarin-4-yl]methyl
(5,7-BCMCM),
[7,8-bis(carboxymethoxy)coumarin-4-yl]methyl
(7,8-BCMCM),
{7-[bis(carboxymethyl)amino]coumarin-4-yl}methyl
(BCMACM),
[6,7-bis(ethoxycarbonylmethoxy)coumarin-4-yl]methyl
(BECMCM),
[7-(diethylamino)-α-methylcoumarin-4-yl]methyl
(DEAMCM),
α-carboxy-4,5-dimethoxy-2-nitrobenzyl (CDMNB),
and 4-carboxymethoxy-2-hydroxycinnamoyl (CMHC)
moieties as novel caging groups. Nearly all these caging
groups bear anionic substituents which confer high
aqueous solubility. Furthermore, the coumarinylmethyl
caging groups absorb at wavelengths >350 nm and therefore
allow uncaging at long-wavelength irradiation.
Using the novel coumarinylmethyl protecting groups and
our earlier introduced caging groups, we prepared improved
caged versions of cyclic nucleoside monophosphates
(cNMPs). Their usefulness was demonstrated in physiological
studies of different types of cyclic nucleotide-gated
ion channels of olfactory sensory neurons (B. Wiesner,
FMP; A. Menini, Trieste; K. Benndorf, Jena) and of sperm
(U. B. Kaupp, Jülich). The most useful caged cNMPs are
the BCMACM, BCMCM and BECMCM-caged derivatives.
We could show that photocleavage of these coumarinylmethyl-caged
compounds was possible with two-photon
excitation as well (collaboration with B. Wiesner, FMP)
and that the liberation of the cNMPs occur in the nanosecond
time scale. A fluorescence spectroscopic method for
the calculation of the rate constants of the steps of the
photolysis pathways was developed (collaboration with J.
Bendig, Berlin and R. Schmidt, Frankfurt/Main). The high
soluble BCMACM-caged compounds allow unprecedented
large instantaneous steps of the cNMP concentration.
Using membrane-permeable BECMCM-caged cGMP and
cAMP, it was demonstrated for the first time that cGMP
promotes the influx of Ca 2+ into sperm of sea urchin or starfish
and it succeeds in combination with caged resact the
identification of the motor response in free swimming
sperm (Kaupp, Jülich).
The CDMNB group was used for caging and uncaging of
the vanilloid receptor agonist capsaicin (collaboration with
S. Frings, Heidelberg). Caged capsaicin is a novel tool for
kinetic examinations of TRPV1 channels in somatosensory
neurons. First applications of CDMNB-caged capsaicin
to noniceptors from rat dorsal root ganglia confirmed that
it can be used as a phototrigger for the activation of noniceptors
in physiological studies.
The group was also concerned with the development of
caged protons. Protons interact with all proteins, which
modulate their structure and their catalytic properties.
Therefore protons trigger events in protein folding/unfold-

ing and play a crucial role in cellular signal transduction.
Kinetic studies of all these processes may be aided by
photoactivatable proton precursors for the generation of
rapid pH jumps. A number of caged protons have been
described, but they have a lot of disadvantages. We synthesized
and characterized caged protons based on
variants of our newly introduced 7-(dimethylamino)coumarinylmethyl
(DMACM) caging group. The novel caged proton
sodium DMACM sulfate exhibits a photorelease rate
constant of at least 5x10 8 s -1, long-wavelength absorption
properties, a high extinction coefficient and a high quantum
yield. Furthermore, DMACM sulfate is very easily soluble
in water. The combination of high photosensitivity and
high solubility allows to induce large pH jumps. We found
also sensitivity of the compound to two-photon photolysis
(collaboration with B. Wiesner, FMP) that should allow true
three-dimensional resolution. The utility of the caged protons
was demonstrated in studies of the H + migration along
lipid bilayers (P. Pohl, S. Keller, FMP).
Group members
Daniel Geißler (Doctoral student)*
Ralf Lechler (Doctoral student)
Nico Kotzur (Student)*
Brigitte Dekowski (Technical assistance)
External funding
Deutsche Forschungsgemeinschaft
„Neue ‚caged’ cyclische Nucleotide – Synthese, Photochemie
und biologische Anwendungen“ (HA 2694/1-2)
Volker Hagen
European Community
„Cyclic nucleotides for the study of cellular events“ (Bio4-
CT98-0034)
Volker Hagen, U. Benjamin Kaupp (Research Center Jülich)
Selected publications (FMP authors in bold)
Kaupp UB, Solzin J, Hildebrand E, Brown JE, Helbig A,
Hagen V, Beyermann M, Pampaloni F, Weyand I (2003) The
signal flow and motor response controling chemotaxis of
sea urchin sperm. Nature Cell Biol 5, 109-117
* part of period reported
FIGURE 1
Large jumps in pH can be achieved upon
flash photolysis of phototriggers for H +
(F = pH-sensitive fluorescence indicator).
Chemical Biology
87

Geißler D, Kresse W, Wiesner B, Bendig J, Kettenmann H,
Hagen V (2003) DMACM-caged adenosine nucleotides:
Ultrafast phototriggers for ATP, ADP and AMP activated by
long-wavelength irradiation. ChemBioChem 4,162-170
Serowy S, Saparov SM, Antonenko YN, Kozlovsky W,
Hagen V, Pohl P (2003) Structural proton diffusion along
lipid bilayers. Biophys J 84, 1031-1037
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W, Pohl
P, Maric K (2003) Cyclic AMP is sufficient for triggering the
exocytic recruitment of aquaporin-2 in renal epithelial
cells. EMBO Rep 4, 88-93
Hagen V, Frings S, Wiesner B, Helm S, Kaupp UB, Bendig
J (2003) (7-Dialkylaminocoumarin-4-yl)methyl-caged compounds
as ultrafast and effective long wavelength phototriggers
of 8-bromo-substituted cyclic nucleotides. Chem-
BioChem 4, 434-442
Matsumoto M, Solzin J, Helbig A, Hagen V, Ueno S,
Kawase O, Maruyama Y, Ogiso M, Godde M, Minakata H,
Kaupp UB, Hoshi M, Weyand I (2003) A sperm-activating
peptide controls a cGMP-signaling pathway in starfish
sperm. Dev Biol 260, 314-324
Collaborations
Photochemistry of caged compounds
J. Bendig and P. Wessig, Institute of Chemistry, Humboldt
University Berlin
Time-resolved fluorescence spectroscopy
R. Schmidt, Institute for Physical and Theoretical Chemistry,
Johann Wolfgang GoetheUniversity, Frankfurt / Main
Studies of cellular signaling using caged compounds and
caged chemoattractants
U. B. Kaupp, Institute for Biological Information Processing,
Research Center Jülich
Caged vanilloid receptor agonists as tools for kinetic
examinations of TRPV1 channels
S. Frings, Department of Molecular Physiology, University
of Heidelberg
Analysis of receptor systems in CNS using caged compounds
H. Kettenmann, Max Delbrück Center for Molecular Medicine,
Berlin-Buch
Controling of gene expression by using caged compounds
S. Cambridge, Max Planck Institute of Neurobiology,
Munich
Caged cNMPs as tools for studies of gating kinetics of
CNG channels
K. Benndorf, Friedrich Schiller University Jena
Studies of the function of CNG channels on isolated olfactory
sensory neurons using caged cNMPs
A. Menini, International School for Advanced Studies,
Trieste, Italy

MEDICINAL CHEMISTRY
Group Leader: Prof. Jörg Rademann
CHEMICAL BIOLOGY WITH SMALL
MOLECULES
Small molecules are powerful biological tools that can be
employed as bioactive protein ligands. They are used to
elucidate and modulate the functions of proteins; moreover,
they have been efficient in the discovery of novel
proteins or the assignment of an observed functions to
specific proteins. In addition, protein ligands are the
potential starting point for drug development efforts.
Small molecule ligands are generally accessible by chemical
synthesis. They can be targeted by classical organic
synthesis methods. However, for the identification of new
biological activities and the subsequent optimization of the
initial compounds, rationally designed collections of molecules,
denominated as chemical libraries, have to be provided.
Library generation poses novel challenges to synthetic
methods including product isolation and analysis.
Therefore, the starting point of our work is the creation of
synthetic and analytical methodology to furnish designed
chemical libraries. For this purpose combinatorial organic
chemistry and solid phase synthesis are applied and
developed. The syntheses in most cases are operated in
parallel with medium throughput.
On the basis of the developed synthetic methods, focused
screening libraries are designed and constructed. Focused
libraries are composed of potential ligands targeting
one protein or a group of target proteins. They are prepared
for subsequent bioassaying in external collaborations
or in-house screens.
Beyond our own synthetic efforts directed at focused
ligand libraries, we are engaged in the set-up of a designed
generic (base) screen library. This compound collection
is created to target proteins without having a focused
library available or with little structural information known.
In order to limit this library to a manageable size, the focus
will be on the detection of fragment-based interactions
with low or medium affinity. The primary hits identified in
an initial screen can be taken as the starting point for a
subsequent chemical refinement.
For the planning of both synthetic and generic screen
libraries, the input of computational chemistry and molecular
modelling tools is essential. Within the FMP we have
organized a medicinal chemistry panel together with the
modelling group and the Screening Unit for the coherent
planning of library composition and hit evaluation.
To strengthen FMP’s activities in the area of chemical biology,
we co-initiated the national collaboration network
“ChemBioNet” and represent the FMP on the board of this
network. The FMP has offered to set up a central screening
library that is to consist of focused libraries, a generic
screen library of limited size, and specialized compound
collections in which synthetic chemists primarily from outside
the FMP can deposit their compounds to make them
accessible for screening. The library concept will be
accompanied by a database to communicate the contents
of libraries together with screening results to external
users.
In the specific projects, the central part of our work over
the last two years was the development of novel polymer
carriers and polymer reagents for focused library production.
Based on polyethyleneimines, a multi-ton industrial
product, very high loaded resins (“ultraresins”) were
developed and used in the synthesis of peptides, heterocycles
and polymer reagents. The new carriers are very
economical in solid phase synthesis as they allow a much
higher yield of products per gram of resin compared to
commercially available carriers. The ultraresin concept
has been extended towards the synthesis and delivery of
dendritic, multivalent peptides to cell targets. Via a reversible
cross-linking of branched polyethyleneimine, the
facile preparation of high molecular weight dendrimers (up
to 60 kDa) was feasible, carrying a controlled number of
copies of small molecules, e.g. peptide chains or peptidomimetics.
These complex constructs were shown to efficiently
deliver peptide ligands across cell membranes via
an endocytotic pathway. Currently the specific biological
activity of such dendritic constructs is under investigation.
The multivalent and dendritic structure of these artificial
macromolecules is expected to render the peptide
ligands less prone to proteolysis and increase their biological
activity through multivalent binding.
For the preparation of glycolipids we have set up a novel
synthetic strategy for library production that was denominated
as “hydrophobically assisted switching phase
(HASP) - synthesis”. This concept allows for a flexible
phase switching, each reaction step can be carried out in
solution or attached to the hydrophobic carrier phase. The
concept has been employed for repetitive glycosylations
yielding oligosaccharides attached to a hydrophobic
anchor. In the next step, a library of immuno-stimulating
rhamnolipids has been prepared. These molecules are
interesting in that they are low molecular weight compounds
which stimulate the innate immune system in a
similar way to bacterial lipopolysaccharides (LPS). Unlike
LPS, these rhamnolipids do not activate the cytokine
Chemical Biology
89

FIGURE 1
Synthetic methods
& analysis
excretion via the Toll-like receptors (Tlr-II and -IV) by a
hitherto unknown mechanism.
The main focus of the synthetic efforts was in the area of
protease inhibitors. Stabilized phosphoranes were found
to be an efficient tool for polymer-supported C-acylation
reactions. With this concept, it was possible to construct
the central element of protease inhibitors, the isosteric
core, directly on the solid support. As a consequence, all
substituents of peptide isoster inhibitors now can be introduced
and varied flexibly. This variability is a new opportunity
especially in respect to the central side chain, the
P1-site, and thus could be employed to investigate the
effect of P1-site variations on specificity and selectivity of
protease inhibitors. In a model study, conducted with the
plasmepsin II, the malaria-linked aspartyl protease of plasmodium
falciparum, we discovered that P1-site mutations
enhanced the affinity of norstatine inhibitors by the factor
of 60 compared to the natural, i.e. substrate-derived side
chain. Further protease projects yielded reversible inhibitors
of the SARS-main protease (in cooperation with
R. Hilgenfeld, Lübeck) and of industrial targets (patent filed
with Boehringer Ingelheim Pharma in May 2004).
Modelliung/
Structural biology
(Focussed)
libraries
Screening
concepts
A future keystone of our research will be the development
of novel screening concepts. In this area we have initiated
a project on primary screen techniques together with
the Screening Unit. Several collaborative projects within
the FMP and on-Campus have been started as well.
Group members
Dr. Samer Al-Gharabli
Dr. Ludmila Perepellichenko*
Dr. Michael Barth
Dr. Syed Tasadaque Ali Shah*
Dr. Steffen Weik
Jörg Bauer (Doctoral student)
Adeeb El-Dashan (Doctoral student)
Viviane Uryga-Polowy (Doctoral student)*
Franziska Meier (Student)*
* part of period reported

External funding
Deutsche Forschungsgemeinschaft
„Reaktive Intermediate in polymeren Gelen, ihre Anwendung
in parallelen Synthesen von Proteaseinhibitoren
sowie deren biochemische Evaluierung“ (Ra895-2)
Jörg Rademann
Deutsche Forschungsgemeinschaft
„Diversitätsorientierte Synthese von Rhamnolipiden“
(Ra895-3)
Jörg Rademann
Deutsche Forschungsgemeinschaft
Teilprojekt im Graduate College „Chemistry in Interphases“
Jörg Rademann
Land Baden-Württemberg
„Reaktionskaskaden“ (Landesforschungsschwerpunkt)
Jörg Rademann
Boehringer Ingelheim Pharma
Jörg Rademann
Merck Biosciences
Jörg Rademann
Selected publications (FMP authors in bold)
Rademann J (2003) Advanced polymer reagents based on
activated reactants and reactive intermediates: Powerful
novel tools in diversity-oriented synthesis. In: Methods in
Enzymology 369, Bunin BA, Morales G (eds), Academic
Press San Diego, 366-390
Bauer J, Rademann J (2003) Trimellitic anhydride linker
(TAL) – highly orthogonal conversions of primary amines
employed in the parallel synthesis of labeled carbohydrate
derivatives (including 5 pages supplemental material).
Tetrahedron Lett 44, 5019 - 5023
Weik S, Rademann J (2003) A Phosphorane as Supported
Acylanion Equivalent. Linker Reagents for Smooth and
Versatile C-C-Coupling Reactions. Angew Chem Int Ed 42,
2491-2494
Barth M, Rademann J (2004) Tailoring Ultraresins based on
the cross-linking of polyethylene imines. Comparative
investigation of the chemical composition, the swelling,
the mobility, the chemical accessibility, and the performance
in solid phase synthesis of very high-loaded resins.
J Comb Chem 6, 340-349
Barth M, Ali Shah T, Rademann J (2004) High Loading
Polymer Reagents based on Polycationic Ultragels. Polymer-Supported
Reductions and Oxidations with Increased
Efficiency. Tetrahedron – Combinatorial Chemistry Symposium
in Print 60, 8703-8709
Rademann J (2004) Organic protein chemistry: drug discovery
through the chemical modification of proteins. Angew
Chem Int Ed 43, 4554-4556
Barth M, Fischer R, Brock R, Rademann J (2005) Reversible
cross-linking of hyperbranched polymers: A strategy for
the combinatorial decoration of multivalent scaffolds.
Angew Chem Int Ed 44, 1560-1563
Bauer J, Rademann J (2005) Hydrophobically assisted
phase synthesis: the flexible combination of solid-phase
and solution-phase reactions employed for oligosaccharide
Preparation. J Am Chem Soc 127, 7296-7297
Collaborations
G. Klebe, University of Marburg
R. Brock, University of Tübingen
R. Hilgenfeld, University of Lübeck
U. Zähringer, K. Brandenburg, H. Heine, FZ Borstel
T. Mayer, MPI Martinsried
Chemical Biology
91

SCREENING UNIT
Group Leader: Dr. Jens Peter von Kries
SCREENING FOR BIOACTIVE SMALL
MOLECULES IN AN ACADEMIC SET UP -
CHEMICAL BIOLOGY INITIATIVE
Beside their medical potential as drugs which cure diseases,
bioactive small molecules may also serve as powerful
tools for analysis of complex biological systems. Systematic
screening of large compound libraries for bioactive
small molecules which serve as molecular „switches“ in
biological networks may become a key technology in
Chemical Biology. Therefore, the “Forschungsinstitut für
Molekulare Pharmakologie“ provides a Screening Unit for
academic research groups which has been functional
since autumn 2004. Furthermore, the institute supplies the
central compound collection of the German Initiative for
Chemical Biology: ChemBioNet.
Biological networks
About 30,000 genes determine the development and stability
of a human organism. They code for proteins of the
protein synthesis machinery, ion channels, receptors,
kinases, adaptor- or scaffolding proteins of cellular communication
pathways, cell adhesion molecules or proteins
of the immune system. Their correct function is maintained
by complex interactions including enzymatic modifications,
like phosphorylation or proteolytic processing. For
instance, adaptor proteins may serve as scaffolds after
modification by phosphorylation which then in turn recruit
enzymes and their corresponding substrates for further
signal transduction.
Comparative genomics has shown that evolution does not
solely correspond to a growing number of genes, but corresponds
to an increase in the complex combination of
regulatory modules in multifunctional proteins or genes.
Perturbation of these complex interaction networks, i.e. by
mutation, viral infection or environmental influences, may
result in diseases like cancer, cardiac disease or diabetes.
Classical methods for analysis
Many proteins have been positioned into pathways using
classical genetic methods, which are based on loss of
function by mutation and on complementation studies to
define for example the position of a given protein in a pathway.
Biochemical in vitro assays measure enzyme activities
or characterize binding domains of proteins. Methods
in modern molecular biology comprise overexpression
experiments of wild-type or mutant proteins in cell culture.
Many of those strategies result in deletion of all functions
of a given protein (antisense) or result in perturba-
tion of many functions, if the overexpressed domain contains
multiple binding sites for other factors. Contribution
of individual interactions cannot be determined from these
experiments. Although some protein functions have been
identified in the past, only a small part of the complex interactions
and functions of the proteins encoded in the
human genome is known as of yet.
Small molecules: new tools for analysis
The functional characterization of the proteins encoded in
the humane genome and those of a growing number of
model organisms present a big challenge for the biosciences.
Chemistry may become a driving force for solution,
as has been shown by recent publications presenting inhibitors
of protein interactions and of enzymatic activities of
proteins. Molecular “switches“ may be generated by chemical
synthesis which modulate the activity of proteins or
genes. The combination of biological and chemical
methods may allow the investigation of the function of proteins
which have, until now, defied functional analysis.
Small bioactive molecules often bind in hydrophobic pockets
of the surface of proteins. This hydrophobic character
also favors crossing of cellular membranes. As small
molecules extend on only small regions of a protein surface,
they potentially inhibit only individual and not all
functions of a protein. Therefore small molecules offer an
ideal tool for identification of individual contribution of
interactions towards function of multiprotein complexes,
i.e. in biological signaling networks. Furthermore, selective
interference with protein binding or enzyme function
allows the comparison with the consequence of loss of
specific functions in human diseases. Moreover, the data
collection and analyses of interactions of small molecules
with proteins provides insights into substrate recognition
mechanisms and will allow more effective de novo design
of agonists or antagonists in the future.
Small molecules and structural biology
Small molecules, which resemble natural substrates of
enzymes (like ATP), but cannot be processed, fix dynamic
structures of enzymes for x-ray analysis of crystals. They
allow the analysis of the active state of an enzyme. This is
an important feature as many enzymes in cancer become
permanent active after mutation. Therefore, this fixed
structure provides an ideal starting point for the design of
inhibitors. Alternatively, this information has been used to
design an enzyme-specific cosubstrate (ATP) which is only
recognized by a specifically mutated enzyme for labeling
of its specific downstream target proteins. In general,
small molecules may help to stabilize protein structures,
which cannot be structurally analyzed in their absence.

FIGURE 1
Screening robots support to screen the 20.000 compounds of the FMP library in less than one reach
Small molecules and protein families
Pharmaceutical companies prefer to inhibit enzyme activities
for drug development.
Inhibition of protein interactions is not a favored project,
because it has been shown to be extremely difficult. One
cause for problems targeting protein interactions are the
extensive interaction surfaces, which cannot be completely
blocked by small molecules. But mutagenesis studies
of many interaction surfaces demonstrate that essential
contact points cluster in a small region of the interaction
surface. This region is defined as a hot spot of interaction.
As substitutions of single amino acid residues in these
regions (point mutants) abrogate the complex formation of
the proteins, small molecules, which bind there, potentially
interfere in the same manner. In protein families these
hot spots show significant variation in amino acid sequence.
Therefore, after modification small molecules may
even discriminate between family members and allow the
targeting of individual interactions. In summary small
molecules present an ideal tool for analysis of the complex
function of the human genome.
Concept of the Screening Unit
Three main strategies of screening are supported:
(1) Screening with moderate “HTS” using standard ELISA
or fluorescence techniques. The screening lab of the
Screening Unit is well equipped with a liquid handling
robot and automatic dispensers, washers and different
readers to support most standard screening techniques.
(2) Parallel to this, a spot synthesis of peptide libraries
derived from the minimal binding domains, containing permutated
peptides is offered. The screening of this peptide
library characterizes the hot spots for interactions
(3). Use of virtual drug screening for the selection of compounds
to be ordered for each project. The protein surfaces
will be analyzed and used for computer-aided
docking of virtual libraries into pockets of hot spots or
catalytic sites.
ChemBioNet: chemistry and screening facilities
for Chemical Biology
The German Initiative for Chemical Biology, named Chem-
BioNet, is currently organizing an academic network combining
the individual screening initiatives of many research
institutions in close contact with industrial partners (for
Chemical Biology
93

detailed information see: www.chembionet.de) for support
of academic screening projects. This screening network
will also build up a central shared compound library at the
FMP and a database of combined bioactivity profiles and
chemical data of compounds (see www.chembionet.de
page “Datenbank”). The network will be supported by chemistry
for the modification of molecules and by the expertise
of the screening centers for the setup of robust
screening assays. The research projects will identify and
use bioactive compounds for functional analysis of biological
networks in development or in disease situations
like cancer, cardiac disease, autoimmune reactions or
others. Beside the established role of small molecules for
identification and characterization of protein functions,
these compounds may serve for the development of new
drugs for interference with human diseases.
Competitor or partner of pharmaceutical companies?
Screening in an academic setup has the advantage that
no obvious disease relevance is a prerequisite for usage of
small molecules for interference and characterization of
biological functions. A specific inhibitor of an individual
protein function is a valuable tool, even when it “only” provides
a molecular switch for characterization of a function,
without any perspective for serving as a new drug. This
independence from commercial aspects allows free collection
of chemical and biological data of bioactive molecules.
These data are not limited to a few types of enzymes
as they include a much broader panel of targets.
Therefore this database may provide important clues
towards the design of new classes of bioactive molecules.
Another advantage of the independence from commercial
aspects is the chance to identify and characterize the
mode of action of potential drugs years before the disease
relevance becomes established. At this special point
the interests of pharmaceutical companies and academic
Screening Units could partially overlap, since academic
institutions do not have the resources for successful drug
development, and pharmaceutical companies do not want
to focus on protein interactions as targets. Pharmaceutical
companies could get rights of privileged usage of
patents and sponsor the academic units with know how or
money for library enlargement. In summary, this partnership
would benefit people suffering from a disease for
which no drug is available yet.
Current academic projects: Mycobacterium
tuberculosis enzyme inhibitor screens
Infection with Mycobacterium tuberculosis, the causative
bacterium of human tuberculosis, results predominantly in
an asymptomatic persistent infection, often referred to as
latency. Infected individuals are at risk over their lifetime
to convert their asymptomatic infection into what is called
reactivation tuberculosis, a disease state both potentially
fatal and highly contagious. The long persistence of the
bacteria in the host during asymptomatic infection in the
face of a robust host immune response poses fundamental
biological questions. The bacteria could be in a nonreplicative,
metabolically and transcriptionally inactive and
dormant state. Recent molecular genetic approaches have
yielded Mycobacterium proteins and lipids important for
virulence and persistence. Hence, it can be argued that
the bacteria manipulate the host immune response to
ensure their persistence.
It is estimated that nearly 2 billion people currently suffer
from latent Mycobacterium tuberculosis infection. Although
the key front-line antituberculosis drugs are effective
in treating individuals with acute tuberculosis, these
drugs are ineffective in eliminating M. tuberculosis during
the persistent stages of latent infection. Consequently,
therapeutics that directly target persistent bacilli are
urgently needed.
The “Structural Proteomics Consortium” selected proteins
from Mycobacterium tuberculosis on the basis of gene
ablation experiments (MPI für Infektionsbiologie, Berlin)
resulting in reduced infectiveness. The structures of
selected proteins are solved at DESY in Hamburg (EMBL
and MPG research groups) and screened for small molecule
inhibitors at the Screening Unit in Berlin and by Combinature
(NMR-screening, Berlin).
Two primary screens using the 3-isopropylmalate dehydrogenase
(IPMDH, enzyme for biosynthesis of branched
chain amino acids) and sterol 14?-demethylase (CYP51)
resulted in identification of inhibitors with IC 50-values in
the micromolar range (Rajesh et al. 2005). Two structural
classes of inhibitors for CYP51 resemble already published
compounds as estriol (IC 50: 100 µM) or 4-phenylimidazole
(IC 50: 1 mM) demonstrating the reliability of this screen be.
One of the CYP51 antagonists inhibits growth of tuberculosis
bacteria in human macrophages (Stefan HE Kaufmann,
MPI-IB, Berlin). Therefore this compound may be used for
development of new drugs against tuberculosis infections.
The novel compound classes identified will be cocrystallized
to analyze substrate recognition mechanisms in the
catalytic site of the enzyme. The IPMDH inhibitors have
been identified in a dual approach. First, a compound libra-

y of about 37,000 compounds was docked into the catalytic
site of the enzyme using the computer program GOLD
and default library screening settings. From 30 virtual highscore
compounds, 10 were selected due to the proposed
number of hydrogen bonds in the site and tested in the
enzyme assay. One compound demonstrated the highest
affinity of all inhibitors identified (IC 50: 35 µM). Second, the
FMP small molecule library of 20,000 compounds was
screened and inhibitors of IPMDH identified (IC 50: 75-250
µM). Cocrystallization experiments of inhibitors with
respective enzymes have been already started. Their
structures serve as an input for ordering related compound
structures to identify inhibitors with higher affinity
for analysis in model systems which measure infectiveness
of treated bacteria. These projects may result in the
identification of small molecules which enable to identify
the biological mechanisms of Mycobacterium tuberculosis
to escape from immune defence. Beside this result, the
compounds could potentially be of use in the development
of drugs against tuberculosis infections in partnership with
pharmaceutical companies.
The Screening Unit is supported by a grant of the BMBF
(0312992J, “Mycobacterium tuberculosis Strukturproteomik
Konsortium”).
Group members
DC Angelika Ehrlich**
Christoph Erdmann (Technical assistance)*
Collaborations
Matthias Willmanns, EMBL, Hamburg
Manfred S. Weiss, EMBL, Hamburg
Hans Bartunik, MPG, Hamburg
W. Birchmeier, MDC, Berlin
* part of period reported
** part-time
Chemical Biology
95

SCIENTIFIC AND TECHNICAL SERVICES

MICRODIALYSIS
Group Leader: Dr. Regina M. Richter
DIRECT MEASUREMENT OF IN VIVO
PROCESSING OF BRAIN NEUROPEPTIDES
USING MICRODIALYSIS
Elevated cerebral levels of a variety of neuropeptides are
believed to be risk factors for the development of diseases
that are related to the stress syndrome, cardiovascular
dysfunction and, ultimately to neurodegenerative processes
such as Alzheimer’s disease (AD). Specifically, excessive
accumulation and deposition of β-amyloid peptides
(Aβ) form senile plaques in AD brains which are considered
to be one hallmark of the disease. Emerging evidence
suggests that Aβ accumulation is not in all AD cases associated
with increased Aβ production. Impaired clearance
of Aβ is, therefore, considered to be a critical factor in the
development of AD pathology. However, the mechanisms
of the in vivo clearance of these critical neuropeptides are
still not understood, and elimination of Aβ from tissues and
plasma by activation of degrading proteases appears to
have considerable therapeutic potential. Thus, the major
focus of research in our laboratory is to understand the
proteolytic mechanism of in vivo Aβ degradation.
For all investigated peptides we identified primary cleavage
sites and involved proteases by using specific protease
inhibitors. The impact of structural parameters on Aβ
clearance was explored in greater detail by using double
D-amino acid replacement sets and N-C inverted peptides.
In addition, other factors such as astrocytes which may
have a direct role in Aβ degradation have been studied. To
address specific questions of peptide processing, we
include transgenic and gene-targeted mouse lines that either
overexpress or carry deletions of genes of interest.
Our experimental tools comprise the use of reverse microdialysis
both to introduce neuropeptides or inhibitors into
the brain of awake rodents, as well as to collect metabolic
fragments. In collaborative work, advanced mass spectrometric
techniques are applied to monitor the clearance
of these neuropeptides close to real-time in dialysates of
a few microliters volume. Further efforts are directed
towards quantitative analysis of the peptide fragments.
Work report: Major objectives of the laboratory
Normally soluble β-amyloid peptides (Aβ), generated by
proteolytic cleavage of the β-amyloid precursor protein
(APP), is a mainly 40-42 amino acid species which may
extend to Aβ residue 43/46 in neuritic plaques (Zhao et al.,
JBC 49, 50647, 2004). Although multiple proteases such as
neutral endopeptidase (NEP; neprilysin), insulin-degrading
enzyme (IDE; insulysin) and angiotensin-converting enzyme
(ACE) have recently been identified to degrade extracellular
Aβ, the proteolytic pathways are less well understood.
In particular, in vivo studies are rare and differ
evidently from in vitro reports (for review Hersh, Curr
Pharm Des 9, 449, 2003). Reverse microdialysis in combination
with advanced mass spectrometric techniques is used
to study the clearance of these neuropetides in the brain
of rodents (Figure 1).
Using these methods, we have recently demonstrated the
pathways of in vivo processing of several Aβ species by
NEP using specific protease inhibitors. Current studies
focus on the interplay of structural parameters and protease
activity as well as on the contribution of other candidate
proteases to the regulation of brain Aβ levels using relevant
protease knockout and overexpressing mouse lines.
Another project explores whether astrocytes could have a
direct role in Aβ degradation either by Aβ-induced activation
or chemotactic migration to Aβ deposits (Wyss-Coray
et al., Nature Med 9: 453-456, 2003). Astrocyte motility was
demonstrated to be controlled by the ryanodine type 3
receptor (RyR3) (Matyash et al., FASEB J 16: 84-86, 2002).
These findings encouraged us to test the hypothesis that
extracelluar Aβ degradation is reduced in RyR3 knockout
mice associated with impaired astrocyte motility (in collaboration
with H. Kettenmann, MDC, Berlin). Finally, studies are
underway directed at understanding the principles of extensive
amyloid deposition in cerebral vessels leading to cerebral
amyloid angiopathy. While chronically reduced microcirculation
is believed to impair the clearance of circulating
Aβ, the reported vasoconstrictor action of circulating Aβ
(Niwa et al., Am. J. Physiol 281, H2417, 2001) is still controversial.
In order to address these questions, we have initiated
collaborative work with M.J. Mulvany, University of
Aarhus, Denmark, to explore the direct contractile effects
of Aβ in microvessels, as well as the possibility that the
resulting hypoperfusion results in accumulation of Aβ.
A second area of research concerns the generation of bioactive
angiotensin peptides from angiotensin I. In particular,
angiotensin (1-7) [Ang-(1-7)] is thought to mimic and
oppose the multiple actions of angiotensin II. Both the role
of substrate availability and the metabolic pathways yielding
to the formation of Ang-(1-7) remain unclear. Using
protease knockout mouse models and protease inhibitors,
we have elucidated the major pathways of Ang-(1-7) formation
including the involved proteases in vivo.
Main objective 1. Impact of structural parameters on Aβ
processing
Little is known about the impact of structural parameters
on the catabolic pathways of Aβ peptides. Therefore, we

% Intensity
% Intensity
Wildtype mouse
100
90
80
70
60
50
40
30
20
10
5.3E+4
0
799.0 1579.4 2359.8 3140.2 3920.6 4701.0
Mass (m/z)
100
90
80
70
60
50
40
30
20
10
NEP knockout mouse
4.6E+4
0
799.0 1579.4 2359.8 3140.2 3920.6 4701.0
Mass (m/z)
FIGURE 1
Cerebral microdialysis in a freely moving
mouse.
FIGURE 2
Comparison of MALDI mass spectra of
Aβ(1-40) peptide fragments obtained from
wildtype (upper) and NEP knockout mice
(bottom). The peptide was infused into
the hippocampus of the animals at a very
low flow rate. Relative signal intensity is
given in arbitary units; m/z = mass-tocharge
ratio.
99 Scientific and Technical Services

tested the hypothesis that both the sequence of amino
acids and the secondary structure may play a substantial
role in the processing of Aβ. Thus, two N-C inverted Aβ
chains, (42-1) and (40-1), and a double D-amino acid replacement
set of Aβ were involved in the paradigm to study
their proteolytic cleavage pattern. The two inverted Aβ
species serve as inactive controls because of lacking neurotoxic
properties in cell cultures (Simmons et al., Mol
Pharmacol 45, 373, 1994). In contrast to regular Aβ, the
inverted Aβ species were cleaved to only a few short Cterminal
fragments at the peptide bond His 27-His 28, Leu 24-
His 25 and Phe 22-Val 23 in the (40-1) chain (Richter & Kraus,
Soc Neurosci Abstr # 409.3, 2003). The cleavage site at His-
His is considered to be a preferred target of IDE, whereas
cleavage sites at Phe-Phe and in the N-terminal region of
the N-C inverted molecule are thought to be caused by
NEP. The lack of cleavage sites caused by NEP suggests
that only IDE but not NEP was involved in the degradation
of Aβ under these experimental conditions.
Systematic double D-amino acid replacement of adjacent
amino acids within the Aβ(1-42) molecule was used to study
the impact of the secondary structure on Aβ clearance. In
contrast to former reports, more recent studies revealed
that double D-amino acid substitution efficiently inhibited
the peptide degradation primarily around the replaced
amino acids. In aCSF, a complete protection of the molecule
against enzymatic degradation indicating a folding of the
molecule was not found. These results are strongly supported
by findings obtained with Aβ dissolved in buffer and
subsequently analyzed with CD and ITC methods
(S. Keller, unpublished data). These results suggest that the
proteolytic clearance of Aβ peptides in vivo depends highly
on both the primary and secondary structure of the molecule
and differs significantly from that ex vivo.
Main objective 2. Proteases involved in Aβ clearance
First, we explored the two principal regulators of Aβ clearance:
NEP and IDE. The postulated primary cleavage site
at positions 33/44 (Gly-Leu) and the N-terminal directed
ladder-like degradation in rat hippocampus were successfully
blocked by the NEP inhibitors phosphoramidon and
thiorphan, suggesting a major role of this particular protease
in Aβ clearance. However, compared to the literature
our in vivo findings indicate additional metabolic pathways
of Aβ. For example, the intermediate fragment
(10-37), identified as the major product of Aβ(1-42) cleavage
by NEP (Iwata et al., Nature Med 6, 143, 2000), was
not detected in our close to real-time experiments. Moreover,
separate metabolic studies revealed a very rapid
degradation of the fragment which does not correspond to
the Aβ(10-37) accumulation in catabolic studies on radiolabeled
Aβ reported by Iwata and collegues. Also, nume-
rous mass spectra displayed an additional cleavage site at
position 34/35 (Leu-Met) attributed to a matrix metalloproteinase-9
(MMP-9) which is believed to prevent plaque
formation (Backstrom et al., J Neurosci 16:7910, 1996).
To study the particular role of IDE in Aβ degradation, we
extended our studies to awake NEP knockout versus wildtype
mice. The in vivo profile of hippocampal Aβ processing
revealed cleavage patterns which match well to in
vitro findings from Mukherjee and colleagues (J Neurosci
20:8745-8749, 2000) in so far as short N-terminal fragments
(1-11 to 1-16) dominated in mass spectra while the C-terminal
region remained intact (Figure 2) (Richter et al., Soc
Neurosci Abstr # 220.6, 2004). These results provide strong
evidence that the major proteases NEP and IDE contribute
to the clearance of Aβ by cleavage of differing peptide
bonds in distinct regions within the peptide chain.
Group members
Antje Muschter (Technical assistance)* **
Christian Wolff (Technical assistance)**
External funding
Schering AG-Ri1; Regina Richter
Selected publications (FMP authors in bold)
Richter RM, Kraus M (2003). Profile and Inhibition of the in
vivo Degradation of Alzheimer’s β-Amyloid Peptides in Rat
Brain. Soc Neurosci Abstr # 409.3
Richter RM, Heyne A, Schuchardt S (2004). Degradation of
Amyloid β-Peptide in the Hippocampus of a NEP knockout
Mouse Model. Soc Neurosci Abstr # 220.6
Collaborations
Hersh, L.B. (Dept. Molec. and Cellular Biochemistry, University
of Kentucky, Lexington, KY, USA)
Degradation of β-amyloid peptides in transgenic and
knockout mouse models
Kettenmann, H. (Dept. Cellular Neuroscience, MDC for
Molecular Medicine, Berlin, Germany)
Clearance of β-amyloid peptides in the brain of RyR3knockout
mice
Mulvany, M.J. (Dept. Pharmacology, Aarhus University,
Aarhus, Denmark)
Cerebral hypoperfusion and beta-amyloid plaque formation
Schuchardt, S. (PLANTON GmbH), Kiel
MALDI-TOF and MALDI-MS/MS analysis of the in vivo
metabolism of neuropeptides
* part of period reported
** part-time

DNA SEQUENCING
Group Leader: Dr. Erhard Klauschenz
The DNA Sequencing Service Group conducts DNA
sequencing for the molecular research groups of the FMP.
It comprises a scientist and a technician (part-time); two
automated sequencers are available (ABI 377 and ABI
310).
At present about 100 samples are processed weekly. The
Department of Signal Transduction/Molecular Medicine
submits about 60% of the samples, the Junior Research
Group Cellular Signal Processing (Vinkemeier) 30%, with
occasional samples from other groups.
In addition to the service function, the group continues to
work on the molecular analysis of the vasopressin V2
receptor genes of patients (and their families) suffering
from congenital nephrogenic diabetes insipidus (NDI).
Group members
Barbara Mohs (Technical assistance)**
** part-time
ADDITIONAL SERVICES
Scientific workshop
Jürgen Mevert
Michael Uschner
Holger Panzer
Helmut Blick
Stefanie Wendt
Safety Officer
Dr. Hans-Ulrich Heyne
Central glassware washing facility
Uwe Hackel †
Animal housing
Dr. Regina M. Richter
Eva Lojek
Petra Göritz
Academic library
Renate Peters
Scientific and Technical Services
101

Visit of the Federal President on Berlin-Buch Campus 2004
Besuch des Bundespräsidenten auf dem Campus Berlin-Buch 2004
PUBLIC RELATIONS AND THE MEDIA
Dr. Björn Maul
The Forschungsinstitut für Molekulare Pharmakologie
(FMP) uses public funds to conduct the major part of its
research into the basis of the effects of substances on the
organism. In order to give the taxpayer, who provides the
funds, some insight into how these resources are utilized,
the FMP presents its work to the general public in an easily
understood form.
In 2003 and 2004 the FMP successfully participated in
various public exhibitions and presentations. Scientists
from the Institute used exhibits of interest to the layperson
to introduce their fields of research at the Science Fair of
the Free University Berlin, at the “Müncher Wissenschaftstage”,
and at the presentation "Window on Science"
held in the arcades at Potsdamer Platz.
The FMP also participated in the exhibition "Swimming lab
(MS Chemie)" held on a ship in summer of the Year of Chemistry,
2003. The ship, which was initiated by the scienceto-public
platform “Wissenschaft im Dialog”, moored in 25
cities along the Rhine River and attracted more than 40,000
visitors.
Hans-Olaf Henkel, President of the Leibniz Association, during the
inauguration of the 900 Mhz spectrometer
Hans-Olaf Henkel, Präsident der Leibniz-Gemeinschaft, zur Einweihung
des 900 MHz-Spectrometers
In April 2004 the FMP, together with ten other scientific
institutions from Berlin and the German Human Genome
Project (DHGP), celebrated the fiftieth anniversary of the
discovery of the DNA helix structure with an exhibition in
the Berlin Museum of Natural History (Naturkundemuseum)
that received striking attention. About 40,000 people
paid a visit to the exhibition, which was the only one of its
kind in all of Germany.
The FMP constantly strives to inspire enthusiasm for
scientific research in young people of school age. Scientists
regularly visit schools, presenting easily understood
lectures to promote discussion with pupils and teaching
staff. In addition, the FMP once a year awards a practical
course in the Life Science Learning Lab of the Berlin-Buch
Campus to a winner of one of the country-wide competitions
in "Youth in Research.“ In 2004, the college student
Sylke Höhne from Chemnitz won the prize and was hosted
by the FMP in Berlin.
In each of the two reporting years, the FMP took the
opportunity to present itself to the general public during
the Berlin Long Night of the Sciences. At this event, more
than 1000 visitors came to isolate their own DNA from their

Window on science, Potsdamer Platz in Berlin: Opening lecture
Schaufenster der Wissenschaft am Potsdamer Platz 2003: Eröffnungsvortrag
PRESSE- UND ÖFFENTLICHKEITSARBEIT
Dr. Björn Maul
Das Forschungsinstitut für Molekulare Pharmakologie
(FMP) betreibt den überwiegenden Teil seiner Forschung
über die Grundlagen der Wirkung von Stoffen auf den
Organismus mit Geldern der öffentlichen Hand. Um dem
Steuerzahler, der die Gelder aufbringt, Einblick zu geben,
wofür diese Mittel verwendet werden, stellt das FMP
seine Arbeit in leicht verständlicher Form der breiten
Öffentlichkeit vor.
In den Jahren 2003 und 2004 beteiligte sich das FMP
erfolgreich an verschiedenen öffentlichen Ausstellungen
und Präsentationen. Wissenschaftler des Instituts stellten
ihre Forschungsgebiete anhand von für Laien eingängigen
Exponaten auf der Science Fair der Freien Universität Berlin,
auf der Präsentation „Schaufenster der Wissenschaft“
in den Potsdamer-Platz-Arkaden und auf den Münchner
Wissenschaftstagen vor.
Das FMP beteiligte sich am Chemieschiff (MS Chemie) im
Sommer des Jahres der Chemie 2003. Diese von der „Wissenschaft
im Dialog” initiierte, schwimmende Ausstellung
lief 25 Städte entlang des Rheins an und wurde von mehr
als 40,000 Menschen besucht.
Exhibition “50th anniversary of the DNA helix structure” in the museum
of Natural History
Ausstellung „50 Jahre Doppelhelix“ im Naturkundemuseum
Im April 2004 beging das FMP zusammen mit zehn weiteren
wissenschaftlichen Einrichtungen aus Berlin und
Brandenburg und dem Deutschen Humangenom-Projekt
(DHGP) den 50. Jahrestag der Entdeckung der DNA-Helix
mit einer Sonderausstellung im Berliner Naturkundemuseum.
Etwa 50,000 Menschen besichtigten die Austellung,
die die einzige ihrer Art in Deutschland war.
Das FMP versucht konsequent, junge Menschen bereits
im Schulalter für die naturwissenschaftliche Forschung zu
begeistern. Regelmäßig besuchen Wissenschaftler Schulen
und stellen sich dort mit leicht verständlichen Vorträgen
der Diskussion mit Schülern und Lehrern. Das Institut
unterstützt den Wettbewerb „Jugend forscht“. Es vergibt
einmal im Jahr einen Praktikumskurs im Gläsernen Labor
des Campus als Preis für einen ausgewählten Landeswettbewerb
der Aktion. In 2004 erhielt die Abiturientin Sylke
Höhne aus Chemnitz den Preis und war eine Woche lang
Gast des FMP.
In jedem der beiden Berichtsjahre nahm das FMP die
Gelegenheit wahr, sich in der Berliner Langen Nacht der
Wissenschaften einer breiten Öffentlichkeit zu präsentieren.
Insgesamt kamen mehr als 1000 Menschen zu dieser
Veranstaltung an das FMP, insbesondere um den Mitmachkurs
„Meine DNA“ zu absolvieren und die eigene
103 Scientific and Technical Services

“My DNA“ – The Long Night of the Sciences 2003 at the FMP
„Meine DNA“ – Lange Nacht der Wissenschaften 2003 am FMP
saliva. This special offer was first made in 2002 and became
a real attraction over the years.
The FMP greeted delegations from Germany and abroad
and introduced the participants – government delegations,
journalists, students and school pupils – to its work and
that of the entire Campus. In collaboration with BBB Campus
Management GmbH, the directorate consultants briefed
the numerous visitors on content and organization.
With the cooperation of Institute scientists, short lectures,
tours of the laboratories and discussions were available.
To give the media the opportunity to report significant
events at the FMP, press releases were researched, written
and issued. The public relations consultant as well as
scientists gave interviews, e.g. for Deutschlandfunk, the
Hessian and Berlin broadcasters HR and RBB, as well as
Radio Eins, Radio Multi-Kulti, Korea TV, and the audio:link
Internet radio station. The FMP has been reported in the
major Berlin daily newspapers. The national as well as the
regional press have published reports on FMP scientists
and their research. FMP consultants and scientists have
made several contributions to brochures, various periodicals,
e.g. for the Leibniz Association, the Berlin Research
Association, the German Human Genome Project and for
Exhibition “Swimming lab“ 2003
Ausstellung auf der MS Chemie 2003
the Berlin Buch Campus. The public relations consultant
supported telecasts like “nano” (3sat), “Quarks & Co.”
(West German Broadcast WDR), and “ZDF-Expeditionen”
(ZDF) as an adviser and by providing broadcasting
material.
The FMP gave independent presentations at the Parliamentary
Evenings of the Leibniz Association in Brussels in
2003 and the Forschungsverbund Berlin e.V. (Berlin
Research Association) in Berlin in 2004. The Institute
brought the research of its scientists to the attention of the
international biotech community with a presentation at the
international convention BIO 2004.
Personnel
Ulrike Lauterjung (Assistant)

Parliamentary Evening of the Forschungsverbund Berlin 2004
Parlamentarischer Abend des Forschungsverbunds Berlin 2004
DNA aus einer Speichelprobe zu isolieren. Dieses besondere
Angebot wurde im Jahr 2002 vom FMP initiiert und
hat sich zu einer echten Attraktion entwickelt.
Das FMP empfing Delegationen aus dem In- und Ausland
und machte die Teilnehmer – Regierungsdelegationen,
Journalisten, Studenten und Schüler – mit seiner und der
Arbeit des gesamten Campus bekannt. Das Direktorat
bereitete, oft in Zusammenarbeit mit der BBB Campusmanagement
GmbH, die zahlreichen Besuche inhaltlich und
organisatorisch vor. Zusammen mit den Wissenschaftlern
des Hauses wurden kurze Vorträge, Führungen durch die
Labore und Gespräche angeboten.
Um den Medien Anlass zur Berichterstattung über wichtige
Ereignisse am FMP zu geben, wurden Pressemitteilungen
recherchiert, verfasst und veröffentlicht. Der Referent
für Öffentlichkeitsarbeit vermittelte und gab Interviews,
z. B. für den Deutschlandfunk, den Hessischen Rundfunk
(HR), den Rundfunk in Berlin-Brandenburg (RBB), Radio
Eins, Radio Multi-Kulti, Korea TV und die audio:link Internet-Radiostation.
Über das FMP und die FMP-Wissenschaftler
und ihre Forschung wurde in der überregionalen
und regionalen Presse berichtet, so auch in Artikeln wich-
Science Fair 2004
Science Fair 2004
tiger Tageszeitungen der Berliner Presselandschaft. Referenten
und Wissenschaftler des FMP verfassten Beiträge
für Broschüren und verschiedene Periodika, z. B. der Leibniz-Gemeinschaft,
des Forschungsverbunds Berlin e. V.,
des Deutschen Humangenomprojekts und des Campus
Berlin-Buch.
Der Referent für Öffentlichkeitsarbeit unterstützte verschiedene
Fernsehpoduktionen, wie „nano” (3sat),
„Quarks & Co.” (Westdeutscher Rundfunk) und „ZDF-
Expeditionen” (ZDF) als Berater und mit audiovisuellem
Material.
Das FMP beteiligte sich mit eigenständigen Präsentationen
an den Parlamentarischen Abenden der Leibniz-
Gemeinschaft in Brüssel 2003 und des Forschungsverbunds
Berlin e. V. in Berlin 2004. Das Institut machte die
internationale Biotech-Community auf die Forschung seiner
Wissenschaftler mit einer Präsentation auf der BIO
2004 aufmerksam.
105 Scientific and Technical Services

ADMINISTRATION
Head: Thomas Ellermann
In addition to the Administrative Director, the FMP administrative
staff comprises six employees (three full-time and
three part-time). After the positive conclusion of the last
training cycle, the FMP has decided to take on a new
office communication trainee.
A main focus of activity of the administration during the
reporting period was the preparation for the conversion
from the cameralistic system of accounting to a cost and
performance accounting system (CPA) for the institute.
This CPA system will first be applied to the 2006 budget
year.
The Joint Federal-State Commission for Education Planning
and Research Promotion demands that the institutions
of the Leibniz Association introduce program budgets
at the latest for the budget year 2006. The basis for this is
the decision of the heads of the federal and state governments
from autumn 1997 on “Securing and Quality of
Research”.
VERWALTUNG
Leiter: Thomas Ellermann
In der Verwaltung des FMP sind neben dem Verwaltungsleiter
sechs Mitarbeiterinnen beschäftigt (drei Vollzeit und
drei Teilzeit) beschäftigt. Nach dem positiven Abschluss
des letzten Ausbildungszyklus hat sich das FMP außerdem
entschlossen, erneut eine Bürokommunikations-Kauffrau
auszubilden.
Ein Tätigkeitsschwerpunkt der Verwaltung im Berichtszeitraum
war die Vorbereitung zur Umstellung von der Fehlbedarfsfinanzierung
zu einer leistungs- und ergebnisorientierten
Finanzierung des Instituts. Die leistungs- und
ergebnisorientierte Finanzierung ist erstmalig auf das
Haushaltsjahr 2006 anzuwenden.
Die Bund-Länder-Kommission für Bildungsplanung und
Forschungsförderung (BLK) fordert für die Einrichtungen
der Leibniz Gemeinschaft die Einführung eines Programmbudgets
spätestens zum Haushaltsjahr 2006. Grundlage ist
der Beschluss der Regierungschefs des Bundes und der
Länder vom Herbst 1997 zur „Sicherung und Qualität der
Forschung“.
The cost and performance accounting system (CPA) with
its two hierarchical levels, which was implemented in 2002
and has been operative since 2003, was the prerequisite
for the introduction of the program budget. In the project
group established by the Berlin Research Association
(Forschungsverbund Berlin e.V., FVB) the FMP assumed in
2004 the role of pilot institute for all institutes of the FVB.
Following the directives of the Joint Federal-State Commission
on “minimum requirements on program budgets”,
a standard procedure for the fiscal part of the program
budget could be developed for the institutes of the FVB.
Initially, with the data from the CPA from the year 2004, a
program budget for the budget year 2006 was prepared.
Parallel to that, a financial plan was created in coordination
with the State of Berlin as funding allocator.
At the request of FMP scientists, an electronic ordering
system was introduced. A prerequisite for that was the
successful implementation of SAP version 4.6. Research
staff members can now submit their purchase requests in
digital form via the intranet of the institute. After verification
of the entered data, data relevant to the transaction is
Die im FMP 2002 implementierte und seit 2003 voll produktive
Kosten- und Leistungsrechnung (KLR) mit ihren zwei
Hierarchieebenen war die Voraussetzung für die Einführung
des Programmbudgets. In der vom Forschungsverbund
Berlin e. V. eingerichteten Projektgruppe hatte das
FMP 2004 die Rolle des Musterinstituts für alle Einrichtungen
des FVB übernommen. Orientiert an den Vorgaben der
BLK zu den „Mindestanforderungen an Programmbudgets“
konnte ein Standardverfahren für den finanzwirtschaftlichen
Teil des Programmbudgets für die Institute
des FVB erarbeitet werden. Zunächst wurde mit Daten der
KLR vom Jahr 2004 ein Programmbudget für das Haushaltsjahr
2006 aufgestellt. Parallel wurde in Abstimmung
mit dem Zuwendungsgeber Land Berlin ein Wirtschaftsplan
erstellt.
Auf Wunsch der Wissenschaftler des FMP wurde die elektronische
Bestellübermittlung eingeführt. Voraussetzung
dafür war die erfolgreiche Implementierung der SAP-Version
4.6. Wissenschaftlich Beschäftigte haben jetzt die
Möglichkeit, ihre Beschaffungsanträge in digitaler Form
im Intranet des Instituts zu hinterlegen. Softwareseitig
können buchungsrelevante Daten nach Prüfung über eine
Schnittstelle in eine „Bestellanforderung“ (BANF) des
SAP-Systems übernommen werden. Der Buchungsvor-

transferred into SAP via an interface, and an SAP purchase
request is created. With this procedure, the administration’s
transaction workflow is separated from the rest of
the organization and cannot be accessed by third parties.
The aim is to avoid error-prone and time-consuming double
entries, e.g. in connection with chemical designations.
Orders usually leave the institute on the same day they
were entered using the intranet. Expendable material is
often delivered within 24 hours of the order by the scientist,
depending on the supplier. Therefore, there is no need
for costly warehousing of expendable material at the FMP.
Since 2002, together with the Max Delbrück Center for
Molecular Medicine, the FMP has begun three building
projects and has in part completed them in the period
covered by the report. In 2003 the new lab building NMR II
opened with 200 m 2 floor space used by the FMP, and in
2004 the 900 MHz NMR spectrometer started operation.
After completion, together with the MDC, of the animal
laboratory building Erwin Negelein House at the end of
2004, the FMP has new animal laboratories with a total of
385 m 2 at its disposal. Organizing an economically sound
gang der Verwaltung ist bei diesem Verfahren immer vom
Zugriff Dritter abgekoppelt. Ziel ist es fehlerträchtige und
zeitaufwendige Doppelerfassungen, beispielsweise der
Chemikalienbezeichnung, zu vermeiden. Bestellungen verlassen
das Institut in der Regel tagesgenau. Verbrauchsmittel
werden, abhängig vom Lieferanten, nicht selten
innerhalb von 24 Stunden nach der Bestellung durch den
Wissenschaftler ausgeliefert. Eine aufwendige Lagerhaltung
von Verbrauchsmitteln ist so am FMP nicht nötig.
Das FMP hat gemeinsam mit dem Max-Delbrück-Center
für Molekulare Medizin seit 2002 drei Baumaßnahmen
begonnen und zum Teil im Berichtszeitraum abgeschlossen.
In dem 2003 neu errichtete Labor-Gebäude „NMR II“
mit 200 m 2 vom FMP genutzter Fläche wurde 2004 das 900
MHz-NMR-Spektrometer in Betrieb genommen. Nach
Abschluss der Baumaßnahme des gemeinsam mit dem
MDC erstellten Tierlaborgebäudes „Erwin-Negelein-
Haus“ Ende 2004 kann das FMP neue Tierlabore mit insgesamt
385 m 2 nutzen. Die Organisation eines wirtschaftlichen
Umgangs mit diesen neuen Ressourcen in den
nächsten Jahren stellt die Verwaltung des FMP vor große
Anforderungen. Der Mittelaufwand für Tierhaltung wird
sich stark erhöhen.
usage of these new resources in the coming years presents
great challenges for the FMP administration. Costs
for laboratory animal care will greatly increase.
The new Medicinal Genomics Research Building is due to
open in 2006. Five work groups with a total of 40 employees
from the FMP will use the 1.094 m 2 of floor space allotted
to the institute. In the new laboratory buildings labs are
being fitted out with extensive technical equipment.
Personnel
Silvia Mauks (Personnel Manager)
Birgit Sperling (General Administration)* **
Christel Otto (General Administration)
Claudia Messing (General Administration)**
Kerstin Brauße (General Administration)**
Gabriele Schumacher (Secretary)
Dana Hausbeck (Trainee)*
Josephine Passow (Trainee)*
Der Neubau „Medizinische Genomforschung“ wird voraussichtlich
in 2006 bezogen werden. Fünf Arbeitsgruppen
mit insgesamt 40 Mitarbeitern aus dem FMP werden den
dem Institut zuzurechnenden Flächenanteil von 1.094 m 2
nutzen. In den neuen Laborgebäuden entstehen Laborflächen
mit aufwendigen technischen Ausstattungen.
* part of period reported
** part-time
107 Scientific and Technical Services

COMPUTER SERVICES
Head: Thomas Jahn
The EDP Service Group is centrally located to support the
scientific work of the Research Groups and the administrative
infrastructure (Administration, Library, etc.) and is
responsible for the following areas:
• Provision of communication channels and resources
• Operation and extension of network components
• Central data storage, backup and interchange, print services
• Standard equipment for PCs (hardware and software
co-ordination)
• Technical support in preparing publications and presentations
The backbone of all communication channels forms an
efficient network infrastructure consisting of flexible,
structured network connections and an active network
(modular layer 2/3 switches).
All EDP services are based on a heterogenous server (Net-
Ware 6 Cluster, LINUX and Windows 2k Server). The usual
Internet services are made available by the EDP Group
COMPUTER-SERVICE
Leiter: Thomas Jahn
Zur Unterstützung der wissenschaftlichen Arbeit der Forschungsgruppen
und der administrativen Infrastruktur
(Verwaltung, Bibliothek u. a.) ist eine Arbeitsgruppe EDV
zentral angesiedelt, die für folgende Bereiche verantwortlich
ist:
• Bereitstellung von Kommunikationswegen und -ressourcen
• Betrieb und Ausbau aller Netzwekkomponenten
• zentrale Datenspeicherung, -sicherung und -austausch,
Printservices
• Grundausstattung PC’s (Hard- und Softwarekoordinierung)
• technische Unterstützung bei Publikationserstellung
und -präsentationen
Das Rückgrat für alle Kommunikationswege bildet eine
leistungsfähige Netzinfrastuktur, bestehend aus einer
flexiblen strukturierten Netzwerkverkabelung und einem
aktiven Netzwerk (modulare Layer 2/3-Switches).
Basis für alle EDV-Dienstleistungen ist ein heterogener
Serverpool (NetWare 6 Cluster, LINUX und Windows 2k
Server).
(e-mail, www (Intranet), ftp-Server). The services www (for
FMP externally) and e-mail (MX-Service) are provided
through cooperation with the BBB. The DFN Association is
the Internet provider for the FMP. The Berlin Research Association
(FV Berlin) operates SAP-R/3 server architecture for
administrative tasks, used on the client side in the FMP. PC
work stations with corresponding peripherals for graphics
and image processing are available in the library and in a
graphics room.
An EDP Commission advises on investments and developmental
perspectives in the IT field.
The next tasks of the Group will be implementing measures
to increase security, server consolidation and the evaluation
of future Thin Client Applications to reduce administrative
overheads.
Personnel
Ingrid Hermann (Software Administration)
Hans-Werner Pisarz (Service Engineer)
Alexander Heyne (Student)**
Björn Schümann (Student)* **
Durch die Arbeitsgruppe EDV werden die üblichen Internetdienste
angeboten (eMail, www (Intranet), ftp-Server).
Durch Kooperation mit der BBB werden die Services www
(für FMP extern) und eMail (MX-Service) bereitgestellt.
Der Internetprovider des FMP ist der DFN Verein. Der FV
Berlin betreibt eine SAP-R/3-Serverarchitektur für Verwaltungsaufgaben
die clientseitig im FMP genutzt wird.
In der Bibliothek und in einem Grafikraum stehen PC-
Arbeitsplätze mit entsprechender Peripherie für die Grafik-
und Bildverarbeitung zur Verfügung.
Eine EDV-Kommission berät über Investitionen und perspektivische
Entwicklungen im IT-Bereich.
Für die nächste Zukunft bereitet die Gruppe Maßnahmen
zur Erhöhung der Security vor, plant Schritte zur Serverkonsilidierung
und evaluiert künftige Thin-Client-Applikationen
zur Senkung des administrativen Aufwandes.
* part of period reported
** part-time

APPENDIX

PEER REVIEWED ARTICLES 2003
ORIGINALARBEITEN 2003
SECTION STRUCTURAL BIOLOGY
Protein Structure
Castellani F, van Rossum BJ, Diehl A, Rehbein K,
Oschkinat H (2003) Determination of solid-state NMR
structures of proteins by means of three-dimensional 15N-
13C-13C dipolar correlation spectroscopy and chemical
shifts analysis. Biochemistry 42, 11476-11483
Chevelkov V, van Rossum BJ, Castellani F, Rehbein K,
Diehl A, Hoh M, Steuernagel S, Engelke F, Oschkinat H,
Reif B (2003) 1H detection in MAS solid-state NMR spectroscopy
of biomacromolecules employing pulsed field
gradients for residue solvent suppression. J Am Chem Soc
125, 7788-7789
Labudde D, Leitner D, Krüger M, Oschkinat H (2003) Prediction
algorithm for amino acid type with its secondary
structure in proteins (PLATONS) using chemical shifts. J
Biomol NMR 25, 41-53
Ladizhansky V, Jaroniec CP, Diehl A, Oschkinat H, Griffin
RG (2003) Measurement of multiple psi torsion angles in
uniformly 13C, 15N-labeled alpha-spectrin SH3 domain
using 3D 15N-13C-13C-15N MAS dipolar-chemical shift
correlation spectroscopy. J Am Chem Soc 125, 6827-6833
Pires JR, Hong X, Brockmann C, Volkmer-Engert R,
Schneider-Mergener J, Oschkinat H, Erdmann R (2003)
The ScPex13p SH3 domain exposes two distinct binding
sites for Pex5p and Pex14p. J Mol Biol 326, 1427-1435
Reif B, van Rossum B, Castellani F, Rehbein K, Diehl A,
Oschkinat H (2003) Characterization of 1H-1H distances in
a uniformly 2H, 15N-labeled SH3 domain by MAS solidstate
NMR spectroscopy.
J Am Chem Soc 125, 1488-1489
Strauss H (2003) A device for facilitating the use of the
French Press. Analytical Biochemistry 321, 276-277
Toepert F, Knaute T, Guffler S, Pires JR, Matzdorf T,
Oschkinat H, Schneider-Mergener J (2003) Combining
SPOT Synthesis and Nature Peptide Ligation to Create
Large Arrays of WW Domains. Angew Chem Int Ed 42,
1136-1140
Van Rossum B, Castellani F, Pauli J, Rehbein K, Hollander
J, de Groot H, Oschkinat H (2003) Assignment of amide
proton signals by combined evaluation of HN, NN and
HNCA MAS-NMR correlation spectra. J Biomol NMR 25,
217-223
FMP-authors in bold.
Zimmermann J, Kühne R, Volkmer-Engert R, Jarchau T,
Walter U, Oschkinat H, Ball LJ (2003) Design of N-substituted
Peptomer Ligands for EVH1 Domains. J Biol Chem
278, 36810-36818
Solution NMR
Hupfer M, Rübe B, Schmieder P (2003) Origin and diagenesis
of polyphosphate in lake sediments: A 31P-NMR
study. Limnol Oceanogr 49, 1-10
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Protein Sci 12, 491-500
Structural Bioinformatics
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Protein Sci 12, 491-500
Molecular Modelling
Freund C, Kühne R, Park S, Thiemke K, Reinherz EL,
Wagner G (2003) Structural investigations of a GYF domain
covalently linked to a proline-rich peptide. J Biomol NMR
27, 143-149
Karges B, Karges W, Mine M, Ludwig L, Kühne R, Milgrom
E, de Roux N (2003) Mutation Ala171Thr stabilizes the
gonadotropin releasing hormone receptor in its inactive
conformation, causing familial hypogonadotropic hypogonadism.
J Clin Endocrinol Metab 88, 1873-1879
Zimmermann J, Kühne R, Volkmer-Engert R, Jarchau T,
Walter U, Oschkinat H, Ball LJ (2003) Design of N-substituted
Peptomer Ligands for EVH1 Domains. J Biol Chem
278, 36810-36818
Solid State NMR
Chevelkov V, van Rossum BJ, Castellani F, Rehbein K,
Diehl A, Hoh M, Steuernagel S, Engelke F, Oschkinat H,
Reif B (2003) 1H detection in MAS solid-state NMR spectroscopy
of biomacromolecules employing pulsed field
gradients for residue solvent suppression. J Am Chem Soc
125, 7788-7789
Narayanan S, Bösl B, Walter S, Reif B (2003) Importance of
low oligomeric weight species for prion propagation in the
yeast prion system Sup35/Hsp104. Proc Natl Acad Sci
U.S.A. 100, 9286-9291

Reif B, Griffin RG (2003) 1H detected 1H, 15N correlation
spectroscopy in rotating solids. J Magn Reson 160, 78-83
Reif B, van Rossum B, Castellani F, Rehbein K, Diehl A,
Oschkinat H (2003) Characterization of 1H-1H distances in
a uniformly 2H, 15N-labeled SH3 domain by MAS solidstate
NMR spectroscopy. J Am Chem Soc 125, 1488-1489
Protein Engineering
Freund C, Kühne R, Park S, Thiemke K, Reinherz EL,
Wagner G (2003) Structural investigations of a GYF domain
covalently linked to a proline-rich peptide. J Biomol NMR
27, 143-149
SECTION CELLULAR SIGNALLING/
MOLECULAR GENETICS
Cellular Signalling
Tamma G, Klussmann E, Procino G, Svelto M, Rosenthal
W, Valenti G (2003) cAMP induced AQP2 translocation is
associated with RhoA inhibition through RhoA phosphorylation
and interaction with RhoGDI. J Cell Sci 116, 1519-
1525
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W,
Pohl P, Maric K (2003) Cyclic AMP is sufficient for triggering
the exocytic recruitment of aquaporin-2 in renal ephithelial
cells. EMBO Rep 4, 88-94
Protein Trafficking
Engelsberg A, Hermosilla R, Karsen U, Schülein R, Dorken
B, Rehm A (2003) The Golgi protein RCAS1 controls cell
surface expression of tumor-associated O-linked glycan
antigens. J Biol Chem 278, 22998-23007
Anchored Signalling
Tamma G, Klussmann E, Procino G, Svelto M, Rosenthal
W, Valenti G (2003) cAMP induced AQP2 translocation is
associated with RhoA inhibition through RhoA phosphorylation
and interaction with RhoGDI. J Cell Sci 116, 1519-
1525
Tamma G, Wiesner B, Furkert J, Hahm D, Oksche A,
Schaefer M, Valenti G, Rosenthal W, Klussmann E (2003)
The prostagladin E2 analogue sulprotone antagonizes
vasopressin-induced antidiuresis through activation of
Rho. J Cell Sci 116, 3285-94
FMP-authors in bold.
Schmitt R, Klussmann E, Kahl T, Ellison DH, Bachmann S
(2003) Renal expression of sodium transporters and aquaporin-2
inhypothyroid rats. Am J Physiol Renal Physiol 284,
1097-104
Storm R, Klussmann E, Geelhaar A, Rosenthal W, Maric K
(2003) Osmolality and solute composition are strong regulators
of AQP2 expression in renal principal cells. Am J
Physiol Renal Physiol 284, 189-98
Cellular Imaging
Geissler D, Kresse W, Wiesner B, Bendig J, Kettenmann
H, Hagen V (2003) DMACM-Caged Adenosine Nucleotides:
Ultrafast Phototriggers for ATP, ADP, and AMP Activated
by Long-Wavelength Irradiation. ChemBiochem 4, 162-170
Hagen V, Frings S, Bendig J, Lorenz D, Wiesner B, Kaupp
UB (2003) Fluoreszenzspektroskopische Quantifizierung
der Freisetzung von cyclischen Nukleotiden aus photoaktivierbaren[Bis(carboxymethoxy)-cumarin-4-yl]methylestern
in Zellen. Angew Chem 114, 3775-3777
Hagen V, Frings S, Wiesner B, Helm S, Kaupp UB, Bendig
J (2003) [7-(Dialkylamino)coumarin-4-yl]methyl-caged
compounds as ultrafast and effective long-wavelength
phototriggers of 8-Bromo-substituted cyclic nucleotides.
ChemBiochem 4, 434-442
Kamp G, Büsselmann G, Jones N, Wiesner B, Lauterwein
J (2003) Energy metabolism and intracellular pH in boar
(Sus scrofa) spermatozoa. Reproduction 126, 517-525
Siems WE, Maul B, Wiesner B, Becker M, Walther T,
Rothe L, Winkler A (2003) Effects of kinins on mammalian
spermatozoa and the impact of peptidolytic enzymes.
Andrologia 35, 44-54
Wiesner B, Roloff B, Fechner K, Slominski A (2003) Intracellular
calcium measurements of single human skin cells
after stimulation with corticotropin-releasing factor and
urocortin using confocal laser scanning microscopy. J Cell
Sci 116, 1261-1268
Meyer T, Marg A, Lemke P, Wiesner B, Vinkemeier U
(2003) DNA binding controls inactivation and nuclear
accumulation of the transcription factor. Stat1 Genes Dev
17, 1992-2005
Appendix
111

Molecular Cell Physiology
Wallukat G, Podlovski S, Nissen ER, Morwinski R, Csonka
C, Tosaki A, Blasig IE (2003) Functional and structural characterization
of anti-beta1-adrenoceptor auto-antibodies
of spontaneously hypertensive rats. Mol Cell Biochem 251,
67-75
Schroeter ML, Abdul-Khaliq H, Fruhauf S, Hohne R, Schick
G, Diefenbacher A, Blasig IE (2003) Serum S100B is increased
during early treatment with antipsychotics and in deficit
schizophrenia. Schizophrenia Res 62, 231-236
Biochemical Neurobiology
Siems WE, Maul B, Wiesner B, Becker M, Walther T,
Rothe L, Winkler A (2003) Effects of kinins on mammalian
spermatozoa and the impact of peptidolytic enzymes.
Andrologia 35, 44-54
Biophysics
Urbánková E, Voltchenko A, Pohl P, Jezek P, Pohl EE (2003)
Transport kinetics of uncoupling proteins, Analysis of
UCP1 reconstituted in planar lipid bilayers. J Biol Chem
278, 32497-32500
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W, Pohl
P, Maric K (2003) Cyclic AMP is sufficient for triggering the
exocytic recruitment of aquaporin-2 in renal ephithelial
cells. EMBO Rep 4, 88-94
Molecular Genetics
Wieczorek G, Steinhoff C, Schulz R, Scheller M, Vingron
M, Ropers HH, Nuber UA (2003) Gene expression profile of
mouse bone marrow stromal cells determined by cDNA
microarray analysis. Cell Tissue Res 311, 227-237
Barmeyer C, Horak I, Zeitz M, Fromm M, Schultzke JD
(2003) The interleukin-2-deficient mouse model. Pathobiology
70, 139-142
Cellular Signal Processing
Chen X, Bhandari R, Vinkemeier U, Van Den Akker F,
Darnell JE Jr, Kuriyan J (2003) A reinterpretation of the
dimerization interface of the N-terminal domains of STATs.
Protein Sci 142, 361-365
Meyer T, Vinkemeier U, Meyer U (2003) Medizinische
Implikationen pharmakogenomischer Behandlungsstrategien.
Ethik in der Medizin 12, 207-209
FMP-authors in bold.
Meyer T, Vinkemeier U, Meyer U (2003) Evidence-based
medicine - Was geht verloren? Ethik in der Medizin 14,
3-10
Meyer T, Marg A, Lemke P, Wiesner B, Vinkemeier U
(2003) DNA binding controls inactivation and nuclear
accumulation of the transcription factor Stat1. Genes Dev
17, 1992-2005
SECTION CHEMICAL BIOLOGY
Peptide Synthesis
Carpino LA, Ionescu D, El-Faham A, Beyermann M, Henklein
P, Hanay C, Wenschuh H, Bienert M (2003) Complex
polyfluoride additives in Fmoc-amino acid fluoride coupling
processes. Enhanced reactivity and avoidance of
stereomutation. Org Lett 5, 975-977
Carpino LA, Imazumi H, El Faham A, Fernando JF, Zhang C,
Lee Y, Foxman BM, Henklein P, Hanay C, Mugge C,
Wenschuh H, Klose J, Beyermann M, Bienert M (2003)
The uronium/guanidinium peptide coupling reagents:
Finally the true uronium salts. Biopolymers 71, 349
Otte L, Wiedemann U, Schlegel B, Pires JR, Beyermann
M, Schmieder P, Krause G, Volkmer-Engert R, Schneider-
Mergener J, Oschkinat H (2003) WW domain sequence
activity relationships identified using ligand recognition
propensities of 42 WW domains. Protein Sci 12, 491-500
Von Eggelkraut-Gottanka R, Klose A, Beck-Sickinger AG,
Beyermann M (2003) Peptide (alpha)thioester formation
using standard Fmoc-chemistry. Tetrahedron Lett 44,
3551-3554
Wissmann R, Bildl W, Oliver D, Beyermann M, Kalbitzer HR,
Bentrop D, Fakler B (2003) Solution structure and function
of the “Tandem inactivation domain” of the neuronal
A-type potassium channel Kv1.4. J Biol Chem 278, 16142-
16150
Kaupp UB, Solzin J, Hildebrandt E, Brown JE, Helbig A,
Hagen V, Beyermann M, Pampaloni F, Weyand I (2003) The
signal flow and motor response controlling chemotaxis of
sea urchin sperm. Nat Cell Biol 5, 109-117

Mass Spectrometry
Bente M, Harder S, Wiesgigl M, Heukeshoven J, Gelhaus
C, Krause E, Clos J, Bruchhaus I (2003) Developmentally
induced changes of the proteome in the protozoan parasite
Leishmania donovani. Proteomics 3, 1811-1829
Czupalla C, Nürnberg B, Krause E (2003) Analysis of class
I phosphoinositide 3-kinase autophosphorylation sites by
mass spectrometry. Rapid Commun Mass Spec 17, 690-
696
Czupalla C, Culo M, Müller EC, Brock C, Reusch HP,
Spicher K, Krause E, Nürnberg B (2003) Identification and
Characterization of the Autophosphorylation Sites of Phosphoinositide
3-Kinase Isoforms Beta and Gamma. J Biol
Chem 278, 11536-11545
Beattie KA, Ressler J, Wiegand C, Krause E, Codd GA,
Steinberg CEW, Pflugmacher S (2003) Comparative effects
and metabolism of two microcystins and nodularin in the
brine shrimp Artemia salina. Aquat Toxicol 62, 219-226
Karlsson K, Sipia V, Krause E, Meriluoto J, Pflugmacher S
(2003) Mass Spectrometric Detection and Quantification
of Nodularin-R in Flounder Livers. Environ Toxicol Chem 18,
284-288
Kraus M, Bienert M, Krause E (2003) Hydrogen exchange
studies on Alzheimer's amyloid-beta peptides by mass
spectrometry using matrix-assisted laser desorption/ionization
and electrospray ionization. Rapid Commun Mass
Spec 17, 222-228
Kleuss C, Krause E (2003) G-alpha-s is palmitoylated at the
N-terminal glycine. EMBO J 22, 826-832
Synthetic Organic Biochemistry
Geissler D, Kresse W, Wiesner B, Bendig J, Kettenmann
H, Hagen V (2003) DMACM-Caged Adenosine Nucleotides:
Ultrafast Phototriggers for ATP, ADP, and AMP Activated
by Long-Wavelength Irradiation. ChemBiochem 4, 162-170
Hagen V, Frings S, Bendig J, Lorenz D, Wiesner B, Kaupp
UB (2003) Fluoreszenzspektroskopische Quantifizierung
der Freisetzung von cyclischen Nukleotiden aus photoaktivierbaren[Bis(carboxymethoxy)-cumarin-4-yl]methylestern
in Zellen. Angew Chem 114, 3775-3777
Hagen V, Frings S, Wiesner B, Helm S, Kaupp UB, Bendig
J (2003) [7-(Dialkylamino)coumarin-4-yl]methyl-caged
compounds as ultrafast and effective long-wavelength
phototriggers of 8-Bromo-substituted cyclic nucleotides.
ChemBiochem 4, 434-442
FMP-authors in bold.
Kaupp UB, Solzin J, Hildebrandt E, Brown JE, Helbig A,
Hagen V, Beyermann M, Pampaloni F, Weyand I (2003) The
signal flow and motor response controlling chemotaxis of
sea urchin sperm. Nat Cell Biol 5, 109-117
Matsumoto M, Solzin J, Helbig A, Hagen V, Ueno S,
Kawase O, Maruyama Y, Ogiso M, Godde M, Minakata H,
Kaupp UB, Hoshi M, Weyand I (2003) A sperm-activating
peptide controls a cGMP-signaling pathway in starfish
sperm. Dev Biol 260, 314-324
Lorenz D, Krylov A, Hahm D, Hagen V, Rosenthal W, Pohl
P, Maric K (2003) Cyclic AMP is sufficient for triggering the
exocytic recruitment of aquaporin-2 in renal ephithelial
cells. EMBO Rep 4, 88-94
Medicinal Chemistry
Rademann J (2003) Advanced polymer reagents based on
activated reactants and reactive intermediates: powerful
novel tools in diversity-oriented synthesis. Method Enzymol
369, 366-390
Weik S, Rademann J (2003) A phosphorane as supported
acylanion equivalent: linker reagents for smooth and versatile
C-C coupling reactions. Angew Chem Int Ed 42, 2491-
2494
Rademann J (2003) Trimellitic anhydride linker (TAL) –
highly orthogonal conversions of primary amines employed
in the parallel synthesis of labeled carbohydrate derivatives.
Tetrahedon Lett 44, 5019-5023
Appendix
113

PEER REVIEWED ARTICLES 2004
ORIGINALARBEITEN 2004
SECTION STRUCTURAL BIOLOGY
Protein Structure
Brockmann C, Leitner D, Labudde D, Diehl A, Sievert V,
Büssow K, Kühne R, Oschkinat H (2004) The solution structure
of the SODD BAG-domain and a model of the SODD-
BAG/HAP70 complex reveal additional SODD subfamilyspecific
electrostatic interactions. FEBS Lett 558, 101-106
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kuhne R, Oschkinat H (2004) The oxozidid subunit
b8 from human complex I adopts a thioredoxin fold.
Structure 12, 1645-1654
Gaiser OJ, Oschkinat H, Heinemann U, Ball LJ (2004) (1),
(13) C and resonance assignments of C-terminal BRCT
domain from human BRCA1. J Biol NMR 30, 221-222
Gaiser OJ, Ball LJ, Schmieder P, Leitner D, Strauss H,
Wahl M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
structure, backbone dynamics, and association behavior
of the C-terminal BRCT domain from the breast cancer-associated
BRCA1. Biochemistry 43, 15983-15995
Kahmann JD, Wecking DA, Putter V, Lowenhaupt K, Kim
YG, Schmieder P, Oschkinat H, Rich A, Schade M (2004)
The solution structure of the N-terminal domain of E3L
shows a tyrosine conformation that may explain its reduced
affinity to Z-DNA in vitro. Proc Natl Acad Sci U.S.A.
101, 2712-2717
Krabben L, van Rossum BJ, Castellani F, Bocharov E,
Schulga AA, Arseniev AS, Weise C, Hucho F, Oschkinat H
(2004) Towards structure determination of neurotoxin II
bound to nicotinic acetylcholine receptor: a solid-state
NMR approach. FEBS Lett 564, 319-324
Mueller U, Bussow K, Diehl A, Bartl FJ, Niesen FH,
Nyarsik L, Heinemann U (2004) Rapid purification and crystal
structure analysis of a small protein carrying two terminal
affinity tags. J Struct Funct Genomics 4, 217-225
Pahlke D, Leitner D, Wiedemann U, Labudde D (2004)
COPS - Cis/trans peptide bond conformation prediction of
amino acids on the basis of secondary structure informations.
Bioinformatics 10, 27-28
Pope BJ, Zierler-Gould KM, Kuhne R, Weeds AG, Ball LJ
(2004) Solution structure of human cofilin: rationalising the
pH sensitivity of actin binding. J Biol Chem 279, 4840-4848
FMP-authors in bold.
Soukenik M, Diehl A, Leidert M, Sievert V, Büssow K,
Leitner D, Labudde D, Ball LJ, Lechner A, Nägler DK,
Oschkinat H (2004) The SEP domain of p47 acts as a reversible
competitive inhibitor of cathepsin L. FEBS Lett 576,
358-362
Schleinkofer K, Wiedemann U, Otte L, Wang T, Krause G,
Oschkinat H (2004) Comparative Structural and Energetic
Analysis of WW Domain-peptide. J Mol Biol 344, 865-881
Waldmann H, Karaguni IM, Carpintero M, Gourzoulidou E,
Herrm C, Brockmann C, Oschkinat H, Muller O (2004) Sulindac-Derived
Ras Pathway Inhibitors Target the Ras-Ra
Interaction and Downstream Effectors in the Ras Pathway.
Angew Chem Int Ed 43, 454-458
Wiedemann U, Boisguerin P, Leben R, Leitner D, Krause
G, Mölling K, Volkmer-Engert R, Oschkinat H (2004) Quantification
of PDZ Domain Specificity, Prediction of Ligand
Affinity and Rational Design of Super-Binding Peptides. J
Mol Biol 343, 703-718
Zierler-Gould KM, Pope B, Weeds AG, Ball LJ (2004) Letter
to the Editor: backbone and sidechain 1H, 13 C and 15N
resonance assignments of human cofilin. J Biol NMR 29,
429-430
Zimmermann J, Jarchau T, Walter U, Oschkinat H, Ball LJ
(2004) 1H, 13C and 15N resonance assignment of the
human Spred2 EVH1 domain. J Biol NMR 29, 435-436
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2
receptor. J Biol Chem 279, 47254-47263
Solution NMR
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kuhne R, Oschkinat H (2004) The oxozidid subunit
B8 from human complex I adopts a thioredoxin fold.
Structure 12, 1645-1654
Gaiser OJ, Ball LJ, Schmieder P, Leitner D, Strauss H,
Wahl M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
structure, backbone dynamics, and association behavior
of the C-terminal BRCT domain from the breast cancer-associated
BRCA1. Biochemistry 43, 15983-15995
Kahmann JD, Wecking DA, Putter V, Lowenhaupt K, Kim
YG, Schmieder P, Oschkinat H, Rich A, Schade M (2004)
The solution structure of the N-terminal domain of E3L
shows a tyrosine conformation that may explain its reduced
affinity to Z-DNA in vitro. Proc Natl Acad Sci U.S.A.
101, 2712-2717

Scheich C, Leitner D, Sievert V, Leidert M, Schlegel B,
Simon B, Letunic K, Bussow K, Diehl A (2004) Fast identification
on folded human protein domains expressed in
E. coli suitable for structural analysis. BMC Structural Biology
4, 4
Structural Bioinformatics
Neumann S, Krause G, Claus M, Paschke R (2004) Structural
Determinants for G-Protein Activation and Selectivity
in the Second Intracellular Loop of the Thyrotropin Receptor.
Endocrinol 46, 477-485
Kaufmann R, Schulze B, Krause G, Mayr LM, Setmacher U,
Henklein P (2004) Proteinase-activated receptors (PARs)
– The PAR3 Neo-N-terminal peptide TFRGAP interacts
with PAR1. Regulatory Peptides, in press
Kleinau G, Jaeschke H, Neumann S, Laettig J, Paschke R,
Krause G (2004) Identification of a novel epitope in the TSH
receptor ectodomain acting as intramolecular signalling
interface. J Biol Chem 279, 51590-51600
Müller SL, Portwich M, Schmidt A, Utepbergenov DI,
Huber O, Blasig IE, Krause G (2004) The tight junction protein
occludin and the adherens junction protein alphacatenin
share a common interaction mechanism. J Biol
Chem 280, 3747-3756
Schmidt A, Utepbergenov DI, Mueller SL, Beyermann M,
Schneider-Mergener J, Krause G, Blasig IE (2004) Occludin
binds to the SH3-hinge-GuK unit of zonula occludens
protein 1: potential mechanism of tight junction regulation.
Cell Mol Life Sci 61, 1354-365
Schleinkofer K, Wiedemann U, Otte L, Wang T, Krause G,
Oschkinat H (2004) Comparative Structural and Energetic
Analysis of WW Domain-peptide. J Mol Biol 344, 865-881
Wiedemann U, Boisguerin P, Leben R, Leitner D, Krause
G, Mölling K, Volkmer-Engert R, Oschkinat H (2004) Quantification
of PDZ Domain Specificity, Prediction of Ligand
Affinity and Rational Design of Super-Binding Peptides. J
Mol Biol 343, 703-718
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2
receptor. J Biol Chem 279, 47254-47263
FMP-authors in bold.
Molecular Modelling
Baumgart S, Lindner Y, Kuhne R, Oberemm A, Wenschuh
H, Krause E (2004) The contributions of specific amino acid
side chains to signal intensities of peptides in matrix-assisted
laser desorption/ionization mass spectrometry. Rapid
Commun Mass Spec 18, 863-868
Brockmann C, Leitner D, Labudde D, Diehl A, Sievert V,
Büssow K, Kühne R, Oschkinat H (2004) The solution structure
of the SODD BAG-domain and a model of the SODD-
BAG/HAP70 complex reveal additional SODD subfamilyspecific
electrostatic interactions. FEBS Lett 558, 101-106
Brockmann C, Diehl A, Rehbein K, Strauss H, Schmieder
P, Korn B, Kuhne R, Oschkinat H (2004) The oxozidid subunit
B8 from human complex I adopts a thioredoxin fold.
Structure 12, 1645-1654
Gaiser OJ, Ball LJ, Schmieder P, Leitner D, Strauss H,
Wahl M, Kühne R, Oschkinat H, Heinemann U (2004) Solution
structure, backbone dynamics, and association behavior
of the C-terminal BRCT domain from the breast cancer-associated
BRCA1. Biochemistry 43, 15983-15995
Pope BJ, Zierler-Gould KM, Kuhne R, Weeds AG, Ball LJ
(2004) Solution structure of human cofilin: rationalising the
pH sensitivity of actin binding. J Biol Chem 279, 4840-4848
Solid State NMR
Chen Z, Reif B (2004) Measurements of residual dipolar
couplings in peptide inhibitors weakly aligned by transient
binding to peptide amyloid fibrils. J Biol NMR 29, 525-530
Chevelkov V, Chen Z, Bermel W, Reif B (2004) Resolution
enhancement in MAS solid-state NMR by application of
13C homonuclear scalar decoupling during acquisition. J
Magn Reson 172, 56-62
Ventura S, Zurdo J, Narayanan S, Parreno M, Mangues R,
Reif B, Chiti F, Giannoni E, Dobson CM, Aviles FX, Serrano
L (2004) Short amino acid stretches can mediate amyloid
formation in globular proteins: the Src homology 3 (SH3)
case. Proc Natl Acad Sci U.S.A. 101, 7258-7263
Protein Engineering
Heuer K, Kofler M, Langdon G, Thiemke K, Freund C (2004)
Structure of a helically extended SH3 domain of the T cell
adapter protein ADAP. Structure 12, 603-610
Kofler M, Heuer K, Zech T, Freund C (2004) Recognition
sequences for the GYF domain reveal a possible splicosomal
function of CD2BP2. J Biol Chem 279, 28292-28297
Appendix
115

SECTION CELLULAR SIGNALLING/MOLE-
CULAR GENETICS
Cellular Signalling
Gregan B, Schäfer M, Rosenthal W, Oksche A (2004) Fluorescence
resonance energy transfer analysis reveals the
existence of endothelin A and endothelin B receptor
homodimers. J Cardiovasc Pharmacol 44, S30-33
Gregan B, Jürgensen J, Papsdorf G, Furkert J, Schaefer
M, Beyermann M, Rosenthal W, Oksche A (2004) Liganddependent
differences in the internalization and intracellular
trafficking of endothelin A and endothelin B receptor
heterodimers. J Biol Chem 279, 27679-27687
Hällbrink M, Oehlke J, Papsdorf G, Bienert M (2004) Uptake
of cell-penetrating peptides is dependent on the peptide-to-cell-ratio
rather than on peptide concentration.
Biochim Biophys Acta 1667, 222-228
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
Hermosilla R, Oueslati M, Donalies U, Schönenberger E,
Krause E, Oksche A, Rosenthal W, Schuelein R (2004)
Disease-causing V2 Vasopressin Receptors are Retained
in Different Compartments of the Early Secretory Pathway.
Traffic 12, 993-1005
Tsunoda AP, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin-1, Nothing but a water channel. J Biol
Chem 279, 11364-11367
Wietfeld D, Heinrich N, Furkert J, Fechner K, Beyermann
M, Bienert M, Berger H (2004) Regulation of the Coupling
to Different G-Proteins of Rat Corticotropin-Releasing Factor
Receptor Type 1 (rCRFR1) in HEK293 Cells. J Biol Chem
279, 38386-38394
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2
receptor. J Biol Chem 279, 47254-47263
FMP-authors in bold.
Protein Trafficking
Droese J, Mokros T, Hermosilla R, Schuelein R, Lipp M,
Hopken UE, Rehm A (2004) HCMV-encoded chemokine
receptor US28 employs multiple routes for internalization.
Biochem Biophys Res Comm 322, 42-49
Hermosilla R, Oueslati M, Donalies U, Schönenberger E,
Krause E, Oksche A, Rosenthal W, Schuelein R (2004)
Disease-causing V2 Vasopressin Receptors are Retained
in Different Compartments of the Early Secretory Pathway.
Traffic 12, 993-1005
Neuschäfer-Rube F, Rehwald M, Hermosilla R, Schülein R,
Rönnstrand L, Püschel G (2004) Identification of different
Ser/Thr residues in the C-terminal domain of the human
EP4 reseptor for agonist-induced phosphorylation, betaarrestin
interaction and sequestration Biochemistry 379,
573-585
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2
receptor. J Biol Chem 279, 47254-47263
Anchored Signalling
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
Cellular Imaging
Geissler D, Antonenko YN, Schmidt R, Keller S, Krylova
OO, Wiesner B, Bendig J, Pohl P, Hagen V (2004) (Coumarin-4-yl)methyl
Esters as Highly Efficient, Ultrafast Phototriggers
for Protons and Their Application to Acidifying
Membrane Surfaces. Angew Chem Int Ed 44, 1195-1198
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665

Mathas S, Lietz A, Anagnostopoulos I, Hummel F, Wiesner
B, Janz M, Jundt F, Hirsch B, Johrens-Leder K, Vornlocher
HP, Bommert K, Stein H, Dorken B (2004) C-FLIP mediates
resistance of Hodgkin/Reed-Sternberg cells to death
receptor-induced apoptosis. J Exp Med 199, 1041-1052
Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B, Berger
H, Bienert M (2004) Enhancement of intracellular concentration
and biological activity of PNA after conjugation
with a cell-penetrating synthetic model peptide. Eur J Biochem
271, 3043-3049
Tsunoda AP, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin-1, Nothing but a water channel. J Biol
Chem 279, 11364-11367
Wüller S, Wiesner B, Löffler A, Furkert J, Krause G,
Hermosilla R, Schaefer M, Schülein R, Rosenthal W,
Oksche A (2004) Pharmacochaperones post-translationally
enhance cell surface expression by increasing conformational
stability of wild-type and mutant vasopressin V2
receptor. J Biol Chem 279, 47254-47263
Molecular Cell Physiology
Haseloff RF, Blasig IE, Bauer HC, Bauer H (2004) In search
of the astrocytic factor(s) modulating blood-brain barrier
function in brain capillary endothelial cells in vitro. Mol Cell
Neurosi, in press
Moser KV, Reindl M, Blasig IE, Humpel C (2004) Brain
capillary endothelial cells proliferate in response to NGF,
express NGF receptors and secrete NGF after inflammation.
Brain Res 1017, 53-60
Müller SL, Portwich M, Schmidt A, Utepbergenov DI,
Huber O, Blasig IE, Krause G (2004) The tight junction protein
occludin and the adherens junction protein alphacatenin
share a common interaction mechanism. J Biol
Chem 280, 3747-3756
Schmidt A, Utepbergenov DI, Mueller SL, Beyermann M,
Schneider-Mergener J, Krause G, Blasig IE (2004) Occludin
binds to the SH3-hinge-GuK unit of zonula occludens
protein 1: potential mechanism of tight junction regulation.
Cell Mol Life Sci 61, 1354-365
Zassler B, Blasig IE, Humpel C (2004) Protein delivery of
caspase-3 induces cell death in malignant C6 glioma and
brain capillary endothelial cells. J Neuro-Oncol 71, 127-134
FMP-authors in bold.
Biochemical Neurobiology
Walter T, Stepan H, Pankow K, Gembard F, Faber R,
Schultheiss HP, Siems WE (2004) Relation of ANP and BNP
to their N-terminal fragments in fetal circulation: evidence
for enhanced neutral endopeotidase activity and resistance
of BNP to neutral endopeptidase in the fetus. BJOG
111, 452-455
Walter T, Stepan H, Pankow K, Becker M, Schultheiss HP,
Siems WE (2004) Biochemical analysis of neutral endopeptidase
activity reveals independent catabolism of atrial
and brain natriuretic peptide. Biol Chem 385, 179-184
Biophysics
Geissler D, Antonenko YN, Schmidt R, Keller S, Krylova
OO, Wiesner B, Bendig J, Pohl P, Hagen V (2004) (Coumarin-4-yl)methyl
Esters as Highly Efficient, Ultrafast Phototriggers
for Protons and Their Application to Acidifying
Membrane Surfaces. Angew Chem Int Ed 44, 1195-1198
Krylova O, Pohl P (2004) Ionophoric Activity of Pluronic
Block Copolymers. Biochemistry 43, 3696-3703
Saparov SM, Pohl P (2004) Beyond the diffusion limit:
Water flow throught the empty bacterial potassium channel.
Proc Natl Acad Sci U.S.A. 101, 4805-4809
Sun J, Pohl EE, Krylova OO, Krause E, Agapov II,
Tonevitzky AG, Pohl P (2004) Membrane destabilization by
ricin. Eur Biophys J 33, 572-579
Tsunoda AP, Wiesner B, Lorenz D, Rosenthal W, Pohl P
(2004) Aquaporin-1, Nothing but a water channel. J Biol
Chem 279, 11364-11367
Molecular Genetics
Barmeyer C, Harren M, Schmitz H, Heinzel-Pleines U,
Mankertz J, Seidler U, Horak I, Wiedenmann B, Fromm M,
Schulzke JD (2004) Mechanisms of diarrhea in the interleukin-2-deficient
mouse model of colonic inflammation.
Am J Physiol 286, 244-252
Terszowski G, Waskow C, Conradt P, Lenze D, Koenigsmann
J, Carstanjen D, Horak I, Rodewald HR (2004) Prospective
Isolation and global gene expression analysis of
the colony-forming urit-erythrocyte (CFU-E). Blood 105,
1937-1945
Zatovicova M, Tarabkova K, Svastova E, Gibadulinova A,
Mucha V, Jakubickova L, Biesova Z, Ortova-Gut M,
Parkkila S, Parkkila AK, Waheed A, Horak I, Pastorek J,
Pastorekova S (2004) Monoclonal antibodies generated in
CA IX-deficient mice recognize different domains of tumorassociated
hypoxia-induced carbonic anhydrase IX. J
Immunol Methods 282, 117-134
Appendix
117

Cytokine Signalling
Rosenbauer F, Wagner K, Zhang P, Knobeloch KP, Iwama
A, Tenen DG (2004) pDP4, a novel glycoprotein secreted by
mature granulocytes, is regulated by transcription factor
PU.1. Blood 103, 4294-4301
Schuh K, Cartwright EJ, Jankevics E, Bundschu K, Liebermann
J, Williams JC, Armesilla AL, Emerson M, Oceandy
D, Knobeloch KP, Neyses L (2004) Plasma membrane Ca2+
ATPase 4 is required for sperm motility and male fertility J
Biol Chem 279, 28220-28226
Van Spriel AB, Puls KL, Sofi M, Pouniotis D, Hochrein H,
Orinska Z, Knobeloch KP, Plebanski M, Wright MD (2004)
A regulatory role for CD37 in T cell proliferation. J Immunol
172, 2953-2961
Molecular Myelopoiesis
Heymann GA, Carstanjen D, Kiesewetter H, Salama A
(2004) Polymorphism of the a4-subunit of VLA-4 integrin
and bone marrow transplantation. Haematologica 89,
882-884
Terszowski G, Waskow C, Conradt P, Lenze D, Koenigsmann
J, Carstanjen D, Horak I, Rodewald HR (2004) Prospective
Isolation and global gene expression analysis of
the colony-forming urit-erythrocyte (CFU-E). Blood 105,
1937-1945
Cellular Signal Processing
Marg A, Shan Y, Meyer T, Meissner T, Brandenburg M,
Vinkemeier U (2004) Nucleocytoplasmic shuttling by
nucleoporins Nup153 and Nup214 and CRM1-dependent
nuclear export control the subcellular distribution of latent
Stat1. J Cell Biol 165, 823-833
Meissner T, Krause E, Vinkemeier U (2004) Ratjadone and
leptomycin B block CRM1-dependent nuclear export by
identical mechanisms. FEBS Lett 576, 27-30
Meissner T, Krause E, Lödige I, Vinkemeier U (2004)
Arginine Methylation of STAT1: A Reassessment. Cell 119,
587-589
Meyer T, Hendry L, Begitt A, John S, Vinkemeier U (2004)
A single residue modulates tyrosine dephosphorylation,
oligomerization, and nuclear accumulation of stat transcription
factors. J Biol Chem 279, 18998-19007
FMP-authors in bold.
SECTION CHEMICAL BIOLOGY
Peptide Synthesis
Cansarek P, Beyermann M, Koch KW (2004) Thermodynamics
of apocalmodulin and nitric oxide synthase II peptide
interaction. FEBS Lett 577, 465-468
Carpino LA, Krause E, Sferdean CD, Bienert M, Beyermann
M (2004) Dramatically enhanced N to O acyl migration
during the trifluoracetic acid-based deprotection step in
solid phase peptide synthesis. Tetrahedon Lett 46, 1361-
1364
Carpino LA, Krause E, Sferdean CD, Schümann M, Fabian
H, Bienert M, Beyermann M (2004) Synthesis of “difficult“
peptide sequences:application of a depsipeptide technique
to the Jung Redemann 10- and 26-mers and the amyloid
peptide Abeta (1-42). Tetrahedron Lett 45, 7519-7523
Gregan B, Jürgensen J, Papsdorf G, Furkert J, Schaefer
M, Beyermann M, Rosenthal W, Oksche A (2004) Liganddependent
differences in the internalization and intracellular
trafficking of endothelin A and endothelin B receptor
heterodimers. J Biol Chem 279, 27679-27687
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
Klemm C, Schöder S, Glückmann M, Beyermann M, Krause
E (2004) Derivatization of phorphorylated peptides with
S- and N-nucleophiles for enhanced ionization efficiency
in matrix-assisted laser desorption/ionization mass spectrometry.
Rapid Commun Mass Spectrom 18, 2697-2705
Klose J, Wendt N, Kubald S, Krause E, Fechner K,
Beyermann M, Bienert M, Rudolph R, Rothemund S (2004)
Hexa-histidin tag position influences disulfide structure
but not binding behaviour of in vitro folded N-terminal
domain of rat corticotropin-releasing factor receptor type
2a. Protein Sci 13, 2470-2475
Schmidt A, Utepbergenov DI, Mueller SL, Beyermann M,
Schneider-Mergener J, Krause G, Blasig IE (2004) Occludin
binds to the SH3-hinge-GuK unit of zonula occludens
protein 1: potential mechanism of tight junction regulation.
Cell Mol Life Sci 61, 1354-365

Wietfeld D, Heinrich N, Furkert J, Fechner K, Beyermann
M, Bienert M, Berger H (2004) Regulation of the Coupling
to Different G-Proteins of Rat Corticotropin-Releasing Factor
Receptor Type 1 (rCRFR1) in HEK293 Cells. J Biol Chem
279, 38386-38394
Peptide Lipid Interaction/Peptide Transport
Dathe M, Nikolenko H, Klose J, Bienert M (2004) Cyclization
increases the antimicrobial activity and selectivity
of tryptophan and arginine containing hexapeptides. Biochemistry
43, 9140-9150
Hällbrink M, Oehlke J, Papsdorf G, Bienert M (2004)
Uptake of cell-penetrating peptides is dependent on the
peptide-to-cell-ratio rather than on peptide concentration.
Biochim Biophys Acta 1667, 222-228
Kerth A, Erbe A, Dathe M, Blume A (2004) Infrared reflection
absorption spectroscopy of amphipathic model peptides
at the air/water interface. Biophys J 86, 3750-3758
Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B, Berger
H, Bienert M (2004) Enhancement of intracellular concentration
and biological activity of PNA after conjugation
with a cell-penetrating synthetic model peptide. Eur J Biochem
271, 3043-3049
Wessolowski A, Bienert M, Dathe D (2004) Antimicrobial
activity of Arginine -& Tryptophan-rich hexapeptides: the
effects of aromatic clusters, D-amino acid substitution and
cyclization. J Pept Res 64, 159-69
Peptide Biochemistry
Oehlke J, Wallukat G, Wolf Y, Ehrlich A, Wiesner B,
Berger H, Bienert M (2004) Enhancement of intracellular
concentration and biological activity of PNA after conjugation
with a cell-penetrating synthetic model peptide. Eur
J Biochem 271, 3043-3049
Wietfeld D, Heinrich N, Furkert J, Fechner K, Beyermann
M, Bienert M, Berger H (2004) Regulation of the Coupling
to Different G-Proteins of Rat Corticotropin-Releasing Factor
Receptor Type 1 (rCRFR1) in HEK293 Cells. J Biol Chem
279, 38386-38394
Mass Spectrometry
Baumgart S, Lindner Y, Kuhne R, Oberemm A, Wenschuh
H, Krause E (2004) The contributions of specific amino acid
side chains to signal intensities of peptides in matrix-assisted
laser desorption/ionization mass spectrometry. Rapid
Commun Mass Spec 18, 863-868
FMP-authors in bold.
Carpino LA, Krause E, Sferdean CD, Schümann M, Fabian
H, Bienert M, Beyermann M (2004) Synthesis of “difficult“
peptide sequences:application of a depsipeptide technique
to the Jung Redemann 10- and 26-mers and the amyloid
peptide Abeta (1-42). Tetrahedron Lett 45, 7519-7523
Henn V, Edemir B, Stefan E, Wiesner B, Lorenz D, Theilig F,
Schmitt R, Vossebein L, Tamma G, Beyermann M, Krause
E, Herberg FW, Valenti G, Bachmann S, Rosenthal W,
Klussmann E (2004) Identification of a novel A-kinase
anchoring protein 18 isoform and evidence for its role in
the vasopressin-induced aquaporin-2 shuttle in renal principal
cells. J Biol Chem 279, 26654-26665
Hermosilla R, Oueslati M, Donalies U, Schönenberger E,
Krause E, Oksche A, Rosenthal W, Schuelein R (2004)
Disease-causing V2 Vasopressin Receptors are Retained
in Different Compartments of the Early Secretory Pathway.
Traffic 12, 993-1005
Klemm C, Schöder S, Glückmann M, Beyermann M, Krause
E (2004) Derivatization of phorphorylated peptides with
S- and N-nucleophiles for enhanced ionization efficiency
in matrix-assisted laser desorption/ionization mass spectrometry.
Rapid Commun Mass Spectrom 18, 2697-2705
Klose J, Wendt N, Kubald S, Krause E, Fechner K, Beyermann
M, Bienert M, Rudolph R, Rothemund S (2004) Hexahistidin
tag position influences disulfide structure but not
binding behaviour of in vitro folded N-terminal domain of
rat corticotropin-releasing factor receptor type 2a. Protein
Sci 13, 2470-2475
Meissner T, Krause E, Lödige I, Vinkemeier U (2004)
Arginine methylation of STAT1: a reassessment. Cell 119,
587-589
Meissner T, Krause E, Vinkemeier U (2004) Ratjadone and
leptomycin B block CRM1-dependent nuclear export by
identical mechanisms. FEBS Lett 576, 27-30
Rettig H, Krause E, Börner HG (2004) Atom transfer radical
polymerization with polypeptide-initiators: a general
approach to block copolymers of sequence-defined polypeptides
and synthetic polymers. Macromol Rapid Commun
25, 1251-1256
Sidorova MV, Molokoedov AS, Az’muko AA, Kydryavtseva
EV, Krause E, Ovchinnikov MV, Bespalova ZD (2004) The
Use of hydrogen Peroxide for Closing Disulfide Bridges in
Peptides. Russ J Bio Chem 2, 101-110
Sun J, Pohl EE, Krylova OO, Krause E, Agapov II, Tonevitzky
AG, Pohl P (2004) Membrane destabilization by ricin. Eur
Biophys J 33, 572-579
Appendix
119

Synthetic Organic Biochemistry
Geissler D, Antonenko YN, Schmidt R, Keller S, Krylova
OO, Wiesner B, Bendig J, Pohl P, Hagen V (2004) (Coumarin-4-yl)methyl
Esters as Highly Efficient, Ultrafast Phototriggers
for Protons and Their Application to Acidifying
Membrane Surfaces. Angew Chem Int Ed 44, 1195-1198
Medicinal Chemistry
Barth M, Rademann J (2004) Tailoring Ultraresins based on
the cross-linking of polyethylene imines. Comperative
investigation of the chemical composition, the swelling,
the mobility, the chemical accessibility, and the performance
in solid phase synthesis of very high loaded resins.
J Comb Chem 6, 340-349
Barth M, Fischer R, Brock R, Rademann J (2004) Reversible
cross-linking of hyperbranched polymers: A strategy
for the combinatorial decoration of multivalent scaffolds.
Angew Chem Int Ed 44, 1560-1563
Rademann J (2004) High Loading Polymer Reagents based
on Polycationic Ultragels. Polymer-Supported Reductions
and Oxidations with Increased Efficiency. Tetrahedon Lett
60, 8703-8709
Rademann J (2004) Organic Protein Chemistry: Drug Discovery
through the Chemical Modification of Proteins.
Angew Chem Int Ed 116, 4654-4656
Smerdka J, Rademann J, Jung G (2004) Polymer-bound
alkyltriazenes for mild racemization-free esterification of
amino acid and peptide derivatives. J Pept Sci 10, 603-611
Sorg G, Thern B, Mader O, Rademann J, Jung G (2004) Progress
in the preparation of peptide aldehydes via polymer
supported IBX oxidation and scavenging by threonyl resin.
J Pept Sci 11, 142-152
Number of original articles published in peer reviewed journals ordered by impact factor
Anzahl der Originalarbeiten geordnet nach Impact-Faktor
Impact factor 1999 2000 2001 2002 2003 2004
< 3 14 17 24 16 23 28
3 - 4.5 13 8 7 11 6 16
4.5 - 7 9 8 10 12 12 22
> 7 16 10 12 19 13 14
total 52 43 53 58 54 80
FMP-authors in bold.

REVIEWS 2003/2004
ÜBERSICHTSARBEITEN 2003/2004
Abdul-Kaliq H, Schubert S, Stoltenburg-Didinger G, Huebler
M, Troitsch D, Wehsack A, Boettcher W, Schwaller B,
Crausaz M, Celio M, Schroeter ML, Blasig IE, Hetzer R,
Lange PE (2003) Release patterns of astrocytic and neuronal
biochemical markers in serum during and after experimental
settings of cardiac surgery. Neurol Neurosci/
Molecular Markers of Brain Damage 21, 141-150
Ball LJ, Kühne R, Schneider-Mergener J, Oschkinat H
(2003) Recognition of proline rich motifs (PRMs) by small
adaptor domains: An important class of protein-protein
interactions in signal transduction. Angew Chem Int Ed 44,
2852-2869
Meyer T, Vinkemeier U (2004) Nucleocytoplasmic shuttling
of STAT transcription factors. Eur J Biochem 271, 4606-
4612
Oehlke J, Lorenz D, Wiesner B, Bienert M (2004) Studies
on the cellular uptake of substance P- and lysine-rich,
KLA-derived model peptides. J Mol Recognit 17, 1-10
Oehme P, Schmidt J, Hinkel U (2004) Vitamin K und seine
Funktionen im Körper. Pharmazie 29-31
Oehme, P (2004) Theoretische und klinische Aspekte der
Sucht. Meine Begegnungen mit der Suchtforschung.
Medizin und Gesellschaft 51, 109-119
Pohl P (2004) Combined transport of water and ions
through membrane channels. Biol Chem 385, 921-926
Rademann, J (2004) Organic protein chemistry: drug discovery
through the chemical modification of proteins.
Angew Chem Int Ed 43, 4554-4556
Schülein R (2004) The early stages of the intracellular
transport of membrane proteins: clinical and pharmacological
implications. Rev Physiol Biochem Pharmacol 151,
45-91
Vinkemeier U (2004) Getting the message across, STAT.
Design principles of a molecular signaling circuit. J Cell
Biol 167, 197-201
FMP-authors in bold.
CONTRIBUTIONS IN MONOGRAPHS
2003/2004
BEITRÄGE ZU SAMMELWERKEN 2003/2004
2003
Haseloff RF, Krause E, Blasig IE
Proteomics of the brain endothelium: Separation of proteins
by two-dimensional gel electrophoresis and identification
by mass spectrometry
In: Methods of Molecular Medicine: The blood-brain barrier
- Biology and research protocols (Ed.: Nag S) 89, 465-
477, Humana Press 2003, Totowa, NJ, USA
Klussmann E
Protein kinase A
In: Online pharmacology reference database, Elsevier Science
Inc 2003, Amsterdam, The Netherlands
Krause E
Proteomics
In: Encyclopedic Reference of Molecular Pharmacology
(Eds.: Rosenthal W, Offermanns S) 766-770, Springer Verlag
2003, Heidelberg, Germany
Krause G
Molecular Modelling
In: Encyclopedic Reference of Molecular Pharmacology
(Eds.: Rosenthal W, Offermann S) 603-609, Springer Verlag
2003, Heidelberg, Germany
Meyer T, Vinkemeier U
JAK/STAT Pathway
In: Encyclopedic Reference of Molecular Pharmacology,
(Eds.: Rosenthal W, Offermanns S) 527-530, Springer Verlag
2003, Heidelberg, Germany
Oksche A
Endothelins
In: Encyclopedic Reference of Molecular Pharmacology
(Eds.: Rosenthal W, Offermanns S) 339-345, Springer Verlag
2003, Heidelberg, Germany
Schmidt A, Utepbergenov DI, Krause G, Blasig IE (2003)
Direct demonstration of the association of the blood-brain
barrier proteins ZO-1 and occludin using surface plasmon
resonance spectroscopy-effect of SIN-1
In: Blood-Spinal Cord and brain barriers in health and
disease (Eds.: Sharma HS, Westmann J), Elsevier Science/
Academic Press 2003, San Diego, USA
Schülein R, Rosenthal W
Protein Trafficking and Quality Control
In: Encyclopedic Reference of Molecular Pharmacology
(Eds: Rosenthal W, Offermanns S) 758-762, Springer Verlag
2003, Heidelberg, Germany
Appendix
121

2004
Bauer HC, Bauer H, Haseloff RF, Blasig IE
The role of glia in the formation and function of the bloodbrain
barrier
In: Neuroglia 2nd Edition (Eds.: Ramson B, Kettenmann H)
325-333, Oxford University Press 2004, Oxford, UK
Freund C
The Gyf Domain
In: Modular Protein Domains (Eds.: Cesareni G, Gimona M,
Sudol M, Yaffe M) 107, Wiley-VHC 2004, Weinheim, Germany
Hagen V
Uncaging and photoconversion/activation
In: Encyclopedic Reference of Genomics and Proteomics
in Molecular Medicine (Eds.: Ganten D, Ruckpaul K) in
press, Springer Verlag 2004, Heidelberg, Germany
Kuehne R, Krause G, Rosenthal W
Entdeckungsstrategien in der Wirkstoffforschung
In: Handbuch der Psychopharmakologie (Eds: Holsboer F,
Gruender G, Benkert O) in press, Springer Verlag 2004, Heidelberg,
Germany
Klussmann E
Protein Kinase A
In: xPharm 1.0 (Eds.: Enna SJ, Bylund DB), Elsevier 2004,
Amsterdam, The Netherlands
Krause E
Mass Spectrometry: MS/MS
In: Encyclopedic Reference of Genomics and Proteomics
in Molecular Medicine (Eds.: Ganten D, Ruckpaul K) in
press, Springer Verlag 2004, Heidelberg, Germany
Oehme, P
Zwischen Wissenschaft und Politik
In: Sitzungsbericht der Leibniz-Sozietät (Ed.: Steiger KP)
67, Trafo-Verlag 2004, Berlin, Germany
Oehme P, Göres E, Rosenthal W, Ganten D
Pharmakologische Institutionen Berlin-Buch und Berlin-
Friedrichsfelde
In: Geschichte und Wirken der pharmakologischen, klinisch-pharmakologischen
und toxikologischen Institute
(Ed.: Philippou A) 698-711, Berenkamp Verlag Innsbruck
2004, Austria
Oksche A, Pohl P, Krause G, Rosenthal W
Molecular biology of diabetes insipidus
In: Encyclopedia of Molecular Cell Biology and Molecular
Medicine (Ed.: Meyers RA) 301-324, Wiley-VCH 2004,
Weinheim, Germany
FMP-authors in bold.
Rademann J
Combinatorial Chemistry - Concepts and Methods for
Tasks of Molecular Optimization
In: Encyclopedic Reference of Molecular Pharmacology
(Eds.: Offermanns S, Rosenthal W) 257-261, Springer Verlag
2004, Heidelberg, Germany
Rademann J
Solid phase synthesis (SPS) and polymer-assisted solution
phase (PASP) synthesis
In: Highlights in Bioorganic Chemistry (Eds.: Wennemers
H, Schmuck C) 290-293, Wiley-VCH 2004, Weinheim, Germany
Rademann J
Novel polymer- and linker reagents employed for the preparation
of protease inhibitor libraries
In: Highlights in Bioorganic Chemistry (Eds.: Wennemers
H, Schmuck C) 277-290, Wiley-VCH 2004, Weinheim, Germany
Rademann J
Inhibition of Proteases
In: Highlights in Bioorganic Chemistry (Eds.: Wennemers
H, Schmuck C) 293-295, Wiley-VCH 2004, Weinheim, Germany
Rosenthal W, Seyberth H
Besonderheiten der Arzneimitteltherapie im Kindesalter
In: Pharmakotherapie/Klinische Pharmakologie, 12th
Edition (Eds.: Lemmer B, Brune K) 507-516, Springer Verlag
2004, Heidelberg and Berlin, Germany
Schmidt A, Utepbergenov DI, Krause G, Blasig IE
Direct demonstration of association between the bloodbarrier
proteins ZO-1 and occludin using surface plasmon
resonance spectroscopy - Effect of SIN-1
In: Blood-Spinal Cord Barriers in Health and Disease (Ed.:
Sharma HS) 11-17, Elsevier Verlag/Academic Press 2004,
Heidelberg, Germany
Zimmermann J
EVH1/WH1 domains
In: Modular Protein Domains (Eds.: Cesareni G, Gimona M,
Sudol M, Yaffe M) 73-102. Wiley-VHC 2004, Weinheim,
Germany
MONOGRAPHS 2003/2004
MONOGRAPHIEN 2003/2004
Offermanns S (Eds.), Rosenthal W
Encyclopedic Reference of Molecular Pharmacology
Springer Verlag 2004, Heidelberg, Germany

MEMBERSHIPS IN EDITORIAL BOARDS
2003/2004
MITGLIEDSCHAFTEN IN EDITORIAL
BOARDS 2003/2004
Bienert, Michael
Journal of Receptors and Signal Transduction
Taylor and Francis, Philadelphia
Member since 2003
Blasig, Ingolf
Glia
Wiley-Liss, New York
Member since 2004
Oschkinat, Hartmut
Journal of Structural and Functional Genomics
Kluwer Academic Publishers, Netherlands
Rademann, Jörg
Journal of Combinatorial Chemistry
American Chemical Society
Member since 2002
Rosenthal, Walter
Journal of Molecular Medicine
Springer Verlag, Heidelberg
Member 2001-2004
Rosenthal, Walter
Handbook of Experimental Pharmacology Series
Springer Verlag, Heidelberg, Germany
INVITED TALKS 2003/2004
EINGELADENE VORTRÄGE 2003/2004
Becker M (2004)
NEP-deficient mice - a new model for the late onset human
obesity?
Peptidase-Workshop, EMC Rotterdam. Rotterdam, The
Netherlands
Beyermann M (2003)
In vitro folding, disulfide pattern, and characterization of
the first extracellular domain of rat corticotropin-releasing
factor receptor
6th German Peptide Symposium. Berlin, Germany
Bienert M (2003)
Cellular Uptake of Model Amphipathic Peptides, Cellular
Transport Strategies for Targeting of Epitopes, Drugs and
Reporter Molecules – Celltarget
Workshop. Budapest, Hungary
Bienert M (2003)
Cellular Uptake of Peptides: Design and Synthesis of Peptide-Derived
Carriers for the Delivery of Biologically Active
Compounds into Cells
Opening lecture - EU-Project QLK3-CT-2002-01989: Target
specific delivery systems for gene therapy based on cellpenetrating
peptides. Stockholm, Sweden
Bienert M (2004)
Design, Synthesis and Characterization of Beta-Sheet-Forming
Peptides: The FBP 28 WW Domain
10th Akabori Conference. Awaji, Japan
Blasig IE (2003)
Phosphorylation and Protein-Protein Interactions
Gordon Research Conference: Barriers of the Brain. Tilton,
USA
Blasig IE (2003)
Interaction of tight junction proteins - Oligomerization of
ZO-1 and recruitment of occludin
CVB 2003. Amarillo, USA
Blasig IE (2003)
Hinge region of the SH3-GuK unit of ZO-1 is regulated by
occludin and oligomerization of ZO-1
6th Symposium Signal Transduction of the Blood-Brain
Barriers. Szeged, Ungarn
Blasig IE (2004)
Struktur, Funktion und Dichtheit der Blut-Hirnschranke
Hahn-Meitner-Institut. Berlin, Germany
Appendix
123

Carstanjen D (2003)
Die Rolle des Interferon induzierten Transkriptionsfaktors,
ICSBP, in der Reifung und Funktion von Zellen des myelopoetischen
Systems
Charitè Universitätsmedizin, Campus Benjamin Franklin.
Berlin, Germany
Dathe M (2004)
Apolipoprotein E-derived peptides and drug delivery to the
brain
7 th Symposium Signal Transduction of the Blood Brain
Barriers. Potsdam, Germany
Freund F (2003)
Molecular recognition of proline-rich sequences by the
GYF domain
Max-Planck-Institut für Biophysikalische Chemie. Göttingen,
Germany
Freund F (2004)
Struktur-Funktionsbeziehungen wichtiger T-Cell-Proteine
BMBF Bundesministerium für Bildung und Forschung.
Berlin, Germany
Freund F (2004)
Proline-rich sequence recognition by GYF domains
Universität des Saarlandes. Saarbrücken, Germany
Freund F (2004)
Molecular Recognition
Max-Planck-Institut für Biochemie. München, Germany
Freund F (2004)
Proline-rich sequence recognition by GYF domains
Johann Wolfgang Goethe-Universität Frankfurt, Germany
Haseloff RE (2003)
Alterations in the protein expression of rat brain capillary
endothelial cells induced by oxidative stress
6th Symposium Signal Transduction of the Blood-Brain
Barriers. Szeged, Ungarn
Keller S (2004)
Can Cell-Penetrating Peptides Cross Lipid Membranes?
Institut für Physikalische Chemie. Martin-Luther-Universität
Halle, Germany
Klussmann E (2004)
Anchored cAMP signalling in the vasopressin-induced
aquaporin-2 shuttle in renal principal cells
The Biotechnology Centre of Oslo. University of Oslo, Norway
Knobeloch KP (2003)
Zytokin-Rezeptoren und Zytokin-abhängige Signalwege:
the role of Interferon Consensus Sequence Binding Protein
in hematopoiesis
International Hannover Workshop on Cytokine Receptors
and Cytokine Signaling. Hannover, Germany
Krause E (2004)
Massenspektrometrie in der Proteomforschung
Bundesanstalt für Materialforschung. Berlin-Adlershof,
Germany
Krause G (2004)
Gemeinsamkeiten von Struktur und Funktion des Tight
Junction Proteins Occludin und des Adherens Junction
Proteins alpha-Catenin
Institut für Chemie und Pathobiochemie der Freien Universität
Berlin, Germany
Krause G (2004)
The tight junction protein occludin and the adherens
junction protein alpha-catenin share a common interaction
mechanism with ZO-1
7th International Symposium Signal Transduction in the
blood-brain barriers. Potsdam, Germany
Krause G (2004)
Identifizierung von Wechselwirkungsepitopen zwischen
Zellkontaktproteinen der tight junctions
Workshop Neue Peptidtechnologien. Max Brünger Zentrum,
Universität Leipzig, Germany
Krause G (2004)
Structural determinants for the activation of glycoprotein
hormon receptors
Institut für Reproduktionsmedizin, Universität Münster,
Germany
von Kries JP (2004)
Screening for beta-Catenin Antagonists
Max-Planck-Institute of Molecular Cell Biology and Genetics
Dresden, Germany
von Kries JP (2004)
Screening in an academic setup
EMBL Hamburg, Germany
Leitner D (2004)
PASTE und PAPST for biomolecules
Spanish NMR User’s meeting. Madrid, Spain
Maul B (2003)
Neuropeptidasen und Alkoholsucht
Zentralinstitut für Seelische Gesundheit. Mannheim, Germany

Oehme P (2004)
Theoretische und klinische Aspekte der Sucht
Zentrum für Human- und Gesundheitswissenschaften, FB
Humanmedizin der Freien Universität Berlin, Germany
Oehme P (2004)
Toleranz als essentieller Schutzmechanismus - Reflektionen
aus pharmakologischer Sicht
Wissenschaftliche Konferenz der Leibniz-Sozietät e.V.
Berlin, Germany
Oschkinat H (2003).
From Gene to Structure as viewed by NMR. Structures of
membrane proteins by solid-state NMR. NMR in Molecular
Biology
Euroconference on Structural Genomics. Obernai, France
Oschkinat H (2003)
NMR-Spectroscopy of membrane proteins
5th International Conference on Molecular Structural Biology.
Vienna, Austria
Oschkinat H (2003)
A concept for structure determination of small membrane
proteins by 3D magic angle spinning NMR and its application
to the x spectrin SH3 domain
45th Rocky Mountain Conference on Analytical Chemistry.
Denver, USA
Oschkinat H (2003)
NMR-Spectroscopy of membrane proteins
8th International Dahlem Symposium on Cellular Signal
Recognition and Transduction. Berlin, Germany
Oschkinat H (2003)
NMR-Spectroscopy of membrane proteins
4th Colloquium on Transport. Rauischholzhausen, Germany
Oschkinat H (2003)
The solid-state MAS-NMR structure of the 62-residue
alpha-spectrin SH3 domain and the potential of NMR for
the investigation of membrane proteins
44th ENC Experimental Nuclear Magnetic Resonance Conference.
Savannah, USA
Oschkinat H (2003)
The solid-state MAS-NMR structure of the 62-residue
alpha-spectrin SH3 domain and the potential of NMR for
the investigation of membrane proteins
The 3rd Alpine Conference on Solid-State Nuclear Magnetic
Resonance. Chamonix-Mont, France.
Oschkinat H (2004)
Protein-protein interaction investigated by solution and
Solid-state NMR
EMBL Heidelberg, Germany
Oschkinat H (2004)
Structure determination of proteins by magic angle spinning
MAS NMR
Barossa Valley, Australia
Oschkinat H (2004)
Protein-protein interactions viewed by solution and solidstate
NMR
Paris, France
Oschkinat H (2004)
Protein-Protein Interactions and Membrane Proteins Investigated
by Solution and Solid-State NMR
University of Florence, Italy
Oschkinat H (2004)
Molecular Profiles - Target search in molecular pharmacology
Max-Delbrück-Center Berlin-Buch, Germany
Oschkinat H (2004)
Protein-protein interactions viewed by solution and solidstate
NMR
Technische Universität München, Germany
Oschkinat H (2004)
Structural genomics, membrane proteins and automation
Ventura, USA
Oschkinat H (2004)
Viewing protein-protein interactions and membrane proteins
by solution and solid-state NMR
Biozentrum der Universität Basel, Switzerland
Oschkinat H (2004)
NMR analysis of protein modules
New York, USA
Oschkinat H (2004)
Structures of membrane proteins by solution an solid and
solid-state NMR
New Jersey, USA
Oschkinat H (2004)
Zelluläre Protein-Interaktion und pharmakologische Interferenz
Charité-Universitätsmedizin Berlin, Germany
Oschkinat H (2004)
Structure determination of proteins by magic angle spinning
MAS NMR
Martin-Luther-Universität Halle-Wittenberg, Germany
Oschkinat H (2004)
NMR approaches to structures of membrane proteins
Ascona, Schweiz
Appendix
125

Oschkinat H (2004)
An apoE-derived peptide mediates uptake of PEG-liposomes
onto brain capillary endothelial cells
Bad-Herrenalb, Germany
Oschkinat H (2004)
NMR-Spectroscopy of membrane proteins
5th International Conference on Molecular Structural Biology.
Vienna, Austria
Oschkinat H (2004)
Quality control of the vasopressin V2 receptor in the ER
and ER/Golgi intermediate compartment
4th Action Meeting on new Drugs and Treatment. Berlin,
Germany
Oschkinat H (2004)
Aufbau der Technologieplattform NMR-Messtechnik für
die Proteomforschung
Braunschweig, Germany
Oschkinat H (2004)
Magic-angel-spinning solid-state NMR of proteins
IFIA-BioNMR. Karlsruhe, Germany
Oschkinat H (2004)
Structure determination of proteins by magic angle spinning
MAS NMR
University of Osaka, Japan
Oschkinat H (2004)
Design, Synthesis and Characterization of ß-Sheet-Forming
peptides: The FBP 28 WW Domain
Awaji, Japan
Oschkinat H (2004)
Protein structure determination by magic-angle spinning
solid state NMR
Lille, France
Oschkinat H (2004)
Protein Structure Determination by Solid State MAS NMR
Frauenchiemsee, Germany
Pankow K (2004)
Influence of the natriuretic peptide structure on degradation
by NEP
Peptidase-Workshop, EMC Rotterdam, The Netherlands
Pohl P (2004)
Fluid transport through membrane channels and epithelial
cells
Tagung der Gesellschaft für Biochemie und Molekularbiologie.
Münster, Germany
Pohl P (2004)
Fluid transport through membrane channels and epithelial
cells
Johannes Keppler Universität, Linzer Winter Workshop.
Linz, Austria
Pohl P (2004)
Fluid transport through membrane channels and epithelial
cells: Calcium and Transport Processes
Universität des Saarlandes in Homburg, Germany
Rademann J (2004)
Polymer-unterstützte C-Acylierungen gegen Malaria
Novabiochem Seminar, Technische Universität Berlin,
Germany
Rademann J (2004)
P1-Variation in Peptidisosteren: Molekulare Diversität
durch die Kombination von C- and N-Acylierungen
Max-Bergmann Kreis. Munster, France
Rademann J (2004)
Molekulare Werkzeuge für die chemische Industrie
Bioclub der Freien Universität Berlin. Berlin, Germany
Rademann J (2004)
P1-Site Diversity in Peptide Isoster Libraries via Smooth
CC-Couplings on Linker Reagents.
ACS National Meeting. Philadelphia, USA
Rademann J (2004)
New linkers, resins and reagents - some problems of polymer-supported
synthesis and attemps to solve them.
ABC Technologies Symposium. Basel, Switzerland
Rademann J (2004)
Molekulare Werkzeuge für die Chemische Biologie
(Antrittsvorlesung)
Freie Universität Berlin, Germany
Rademann J (2004)
New roads to Chemical Diversity: From P1-Site Mutants in
Malaria Inhibitors to Combinatorial Synthesis of Complex
Modified Dendrimers
Quaid-iAzzam University. Islamabad, Pakistan
Rademann J (2004)
Diversitäts-orientierte Synthese von Glycolipiden durch
hydrophob unterstützte Synthese von Glycopeptiden
Forschungszentrum Borstel, Germany
Rademann J (2004)
Reversibly cross-linked dendrimers - A strategy for the
facile synthesis and combinatorial variation of complex,
multivalent protein-mimics
7 th International Symposium on Biomolecular Chemistry -
ISBOC-7. Sheffield, United Kingdom

Reif B (2003)
Molecular Interactions between the yeast prion protein
Sup35 and the molecular chaperone Hsp104
Euresco Conference on NMR in Molecular Biology. Obernai,
France
Reif B (2003)
Aggregation behaviour of misfolding peptides and proteins
modulated by molecular chaperones: solution and solidstate
NMR experiments
IMB-Symposium „Protein Folding and Aggregation – From
Principles to Pathological Implications”. Jena, Germany
Reif B (2003)
Use of Protons in MAS Solid-State NMR in perdeuterated
peptides and proteins
EMBO-ILL workshop on Deuterium labeling techniqes for
biomolecular NMR and neutron scattering. Grenoble,
France
Reif B (2004)
Molecular Interactions between Sup35 and Hsp104 characterized
by Solution State NMR
Baltimore, USA
Reif B (2004)
Use of Protons in MAS Solid-State NMR in perdeuterated
peptides and proteins
Asilomar Conference Center, Pacific Grove, California,
USA
Reif B (2004)
Protons in MAS Solid-State NMR
Forschungszentrum Karlsruhe, Germany
Reif B (2004)
Use of perdeuteration in MAS Solid State NMR
Denver, Colorado, USA
Reif B (2004)
Multidimensional NMR in Structural Biology
Il Ciocco, Italy
Rosenthal W (2003)
Cyclic AMP-triggered exocytosis
VIIIth International Dahlem Symposium on Cellular Signal
Recognition and Transduction. Berlin, Germany
Rosenthal W (2003)
Biomedical research and biotechnology in Berlin-Buch:
Bioprofile – wohin führt der Weg? Von der Heilpflanze zum
Drug Design
BioTechnica - Workshop: International Biotechnology -
New Concepts in Molecular Medicine from Finland and
Germany. Berlin, Germany
Rosenthal W (2003)
Leuchttürme der Berliner Wissenschaft. Biotechnologie in
Berlin-Brandenburg
Veranstaltung der Initiative „An Morgen denken“. Berlin,
Germany
Rosenthal W (2003)
Mechanismen der Antidiurese
Symposium Fortschritt und Praxis der Endokrinologie zur
Eröffnung des ambulanten Hochschulzentrums für Endokrinologie,
Diabetologie und Ernährung. Berlin / Potsdam,
Germany
Rosenthal W (2004)
The human V2 vasopressin receptor: gene, the protein, the
mutation and rescue strategies of mutant receptors
Annual congress of the European Renal Association –
European Dialysis and Transplant Association. Lisbon, Portugal
Schmieder P (2004)
Towards the structure of the chromophore in bacterial
phytochromes
Herbsttagung des Hahn-Meitner-Instituts Berlin, Germany
Schmieder P (2004)
Aspekte der Strukturbestimmung von Proteinen mittels der
NMR-Spektroskopie
Hahn-Meitner-Institut Berlin, Germany
Schülein R (2004)
Compartmentalization of NDI-casing vasopressin V2
receptor mutants in the early secretory pathway
5th Global NDI Conference. Phoenix, USA
Schülein R (2004)
Functional significance of the cleavable signal peptides of
corticotropin-releasing factor receptors
Symposium on Inflammation and Pain. Berlin, Germany
Schülein R (2004)
Intracellular transport of G protein-coupled receptors
along the secretory pathway
Symposium on Neuro-Immune Interaction in Pain. Berlin,
Germany
Siems, WE
ACE and NEP - relations to alcohol preference
Peptidase-Workshop, EMC-Rotterdam, The Netherlands
Sun X (2004)
Development and Properties of domain-selective murine
ACE forms
Peptidase-Workshop, EMC-Rotterdam, The Netherlands
Appendix
127

EXTERNAL FUNDING 2003/2004
DRITTMITTEL
7000
6000
5000
4000
3000
2000
1000
0
1583
2105
2178
6867
1998 1999 2000 2001 2002 2003 2004
Others 11 3 6 3 5 37 28
Foundations 216 174 112 2 231 158 116
Industry 92 53 29 51 134 197 110
EU 43 74 217 122 710 414 1984
Berlin 52 54 51 0 0 0 0
BMBF 514 696 773 5422 1045 974 886
DFG 656 1050 992 1267 1073 1383 1349
2989
Distribution of annual expenditure shown for the sources of income since 1998
Verteilung der jährlichen Ausgaben über die Drittmittelgeber seit 1998
(total expenses per year in italics)
(kursiv: Gesamtausgaben pro Jahr)
3163
4473

T€
T€
1500
1250
1000
750
500
250
0
2000
1500
1000
500
0
1383
974
0
2003
DFG BMBF Berlin EU Industry Foundations Others
1349
886
Expenses during the period reported: contribution of the sources of income
Drittmittelausgaben im Berichtszeitraum: Verteilung über die Drittmittelgeber
0
414
2004
1984
DFG BMBF Berlin EU Industry Foundations Others
197
110
158
116
37
28
External Funding
129

PARTICIPATION IN RESEARCH NETWORKS
2003/2004
BETEILIGUNG AN NETZWERKEN UND
VERBUNDPROJEKTEN 2003/2004
Sonderforschungsbereiche der Deutschen
Forschungsgemeinschaft
Sonderforschungsbereich 449: Struktur und Funktion
membranständiger Rezeptoren
Teilprojekt A3: Struktur und Funktion von Transportsignalen
des Vasopressin V2-Rezeptors
Ralf Schülein, Walter Rosenthal
Laufzeit: 01.99-12.04
Sonderforschungsbereich 449: Struktur und Funktion
membranständiger Rezeptoren
Teilprojekt B1: Bestimmung der Raumstrukturen von
Rezeptor-gebundenen Agonisten und Antagonisten mittels
Festkörper-NMR-Spektroskopie
Hartmut Oschkinat
Laufzeit: 01.02-12.04
Sonderforschungsbereich 449: Struktur und Funktion
membranständiger Rezeptoren
Teilprojekt A6: Untersuchungen von CRF- und CRF-Rezeptor-Mutanten
zur Entwicklung eines Modells für die
Liganderkennung von G-Protein-gekoppelten Rezeptoren
der Familie 2
Michael Bienert, Michael Beyermann, Walter Rosenthal
Laufzeit: 01.99-12.04
Sonderforschungsbereich 498: Protein-Kofaktor-Wechselwirkungen
in biologischen Prozessen
Teilprojekt C1: Schlüsselreaktionen der biologischen Wasserstoffaktivierung
am Beispiel der [NiFe]-Hydrogenasen
Hartmut Oschkinat, Bärbel Friedrich (HU)
Laufzeit: 01.03-12.05
Sonderforschungsbereich 498: Protein-Kofaktor-Wechselwirkungen
in biologischen Prozessen
Teilprojekt B6: NMR-spektroskopische Untersuchungen
von lichtinduzierten Strukturveränderungen in Protein-
Chromophorkomplexen
Peter Schmieder
Laufzeit: 01.03-12.05
Sonderforschungsbereich 366: Signalerkennung und
-Umsetzung
Teilprojekt Z3: Herstellung und Haltung genetisch veränderter
Mäuse
Elvira Rohde, Ivan Horak
Laufzeit: 01.09-12.05
Sonderforschungsbereich 366: Signalerkennung und
-Umsetzung
Teilprojekt A11: Verbreitung und Bedeutung N-terminaler
Signalpeptide bei G-Protein-gekoppelten Rezeptoren
Ralf Schülein, Walter Rosenthal
Laufzeit: 01.97-12.05
Sonderforschungsbereich 594: Molekulare Maschinen in
Proteinfaltung und Proteintransport
Teilprojekt A3: Biochemische und NMR-Strukturuntersuchungen
an Sup35p im Komplex mit Hsp104, Hsp40 und
Hsp70
Bernd Reif
Laufzeit 05.01-04.04
Forschergruppen der Deutschen
Forschungsgemeinschaft
Forschergruppe 299: Optimierte molekulare Bibliotheken
zum Studium biologischer Erkennungsprozesse
Teilprojekt 2/2-1: Struktur, Stabilität und Spezifikation von
nichtkatalytischen Proteindomänen und deren Verwendung
als Werkzeuge für das Design einer stabilen minimalen
ß-Faltblattstruktur und das Verständnis von pathologischen
Prozessen
Hartmut Oschkinat, Michael Bienert
Laufzeit: 06.01-12.05
Forschergruppe 299: Optimierte molekulare Bibliotheken
zum Studium biologischer Erkennungsprozesse
Teilprojekt 2/2-2: Theoriegestützte NMR-spektroskopische
Analyse von Protein-Ligand-Wechselwirkungen unter Verwendung
von Peptid-Bibliotheken
Hartmut Oschkinat
Laufzeit: 06.01-12.05
Forschergruppe 299: Optimierte molekulare Bibliotheken
zum Studium biologischer Erkennungsprozesse
Teilprojekt 7/2-1: Studium der Ligand-Erkennung von
CRH-Rezeptoren mit Peptid- und nichtpeptidischen Bibliotheken
Michael Bienert, Jens Schneider-Mergener
Laufzeit: 06.99-12.05
Forschergruppe 463: Innovative Arzneistoffe und Trägersysteme
– Integrative Optimierung zur Behandlung entzündlicher
und hyperproliferativer Erkrankungen
Teilprojekt 7B: Hirntargeting mittels oberflächenmodifizierter
Nanosuspensionen und Apolipoprotein E-Peptid
beladener Trägersysteme
Margitta Dathe
Laufzeit: 11.01-12.07

Graduiertenkollegs der Deutschen Forschungsgemeinschaft
GK 238/3: Schadensmechanismen im Nervensystem – Einsatz
von bildgebenden Verfahren
Ingolf Blasig
Laufzeit: 01.03-12.05
GK 276: Signalerkennung und -umsetzung
Alexander Oksche, Walter Rosenthal
Laufzeit: 10.99-09.05
GK 865: Vaskuläre Regulationsmechanismen
Alexander Oksche, Walter Rosenthal
Laufzeit: 04.03-03.07
GK 441: Chemie in Interphasen
Jörg Rademann
Laufzeit: 10.99-09.07
Leitprojekte des Bundesministeriums für
Bildung und Forschung
Proteinstrukturfabrik 01 GG 9812: Strukturanalyse mit
hohem Durchsatz für medizinisch relevante Proteine
Teilprojekt 9: NMR-Spectroscopy
Hartmut Oschkinat, Peter Schmieder, Dietmar Leitner
Laufzeit: 10.98-09.04
Proteinstrukturfabrik 01 GG 9812: Strukturanalyse mit
hohem Durchsatz für medizinisch relevante Proteine
Teilprojekt 10: NMR-structure determination
Hartmut Oschkinat, Dirk Labudde, Ilia Poliakov
Laufzeit: 10.98-09.04
Verbundprojekte des Bundesministeriums für
Bildung und Forschung
Verbundprojekt 0312992J: Proteomische Methoden für
molekulare Strukturen von Zielproteinen aus dem Mycobacterium
tuberculosis Genom und ihrer Ligandkomplexe
zur Suche von Wirkstoffen
Hartmut Oschkinat, Jens von Kries
Laufzeit: 04.04-06.06
Verbundprojekt 0312890G: Proteomweite Analyse membrangebundener
Proteine
Hartmut Oschkinat
Laufzeit: 04.03-03.06
EU-Projekte
EU-Projekt QLK3-2000-00924: Exploiting synthetic SH2scaffolded
repertoire libraries to profile cancer cells and
interfere with cancer-related phenotypes
Hartmut Oschkinat
Laufzeit: 12.00-30.11.03
EU-Projekt QLRT-2000-00987: Antidiuretics using shortaction
vasopressin – V2-receptor agonists as a new therapeutic
strategy of urinary incontinence and voiting disorders
Walter Rosenthal
Laufzeit: 09.01-08.04
EU-Projekt QLK3-CT-2002-02149: Anchored cAMP signalling
– Implications for treatment of human disease
Enno Klussmann, Walter Rosenthal
Laufzeit: 11.02-20.05
EU-Projekt QLK3-CT-2002-01989: Target specific delivery
systems for gene therapy based on cell-penetrating peptides
Michael Bienert, Johannes Oehlke, Margitta Dathe
Laufzeit: 03.03-02.06
Appendix
131

COOPERATIONS WITH CONTRACT
2003/2004
VERTRAGLICHE KOOPERATIONEN 2003/2004
Vereinbarung über die Zusammenarbeit zwischen der
Freien Universität Berlin und dem Forschungsverbund
Berlin e. V. für das FMP
- Freie Universität Berlin
Vereinbarung über die Zusammenarbeit zwischen der
Charité – Universitätsmedizin Berlin und dem Forschungsverbund
Berlin e. V. für das FMP
- Charité – Universitätsmedizin Berlin
Kooperationsvereinbarung: Gemeinsamer Neubau und
Nutzung des Genomzentrums
- Max-Delbrück-Centrum für Molekulare Medizin, Berlin,
Germany
Kooperationsverbarung: Gemeinsame Nutzung des Hermann-von-Helmholtz-Hauses
- Max-Delbrück-Centrum für Molekulare Medizin, Berlin,
Germany
Kooperationsvereinbarung für das Verbundprojekt „Proteomweite
Analyse membrangebundener Proteine (Pro-
Amp)
- Johann Wolfgang Goethe-Universität Frankfurt, Germany
- GSF-Forschungszentrum für Umwelt und Gesundheit
GmbH, Germany
- Max-Planck-Gesellschaft zur Förderung der Wissenschaften
e. V., Germany
Kooperationsvereinbarung: Ausbau und Intensivierung
der wissenschaftlichen Zusammenarbeit
- Deutsches Primatenzentrum GmbH, Göttingen, Germany
- Bernhard-Nocht-Institut für Tropenmedizin, Hamburg,
Germany
- Heinrich-Pette-Institut für experimentelle Virologie und
Immunologie, Hamburg, Germany
- Forschungszentrum Borstel, Zentrum für Medizin und
Biowissenschaften, Borstel, Germany
- Institut für Molekulare Biotechnologie e.V., Jena
Kooperationsvereinbarung: Automatisierte Methoden für
molekulare Strukturen von biologischen Makromolekülen
aus dem Mycobakterium-tuberculosis-Genom und
ihrer Ligandkomplexe zur Suche von Wirkstoffen mit
Hochdurchsatz Chiptechnologien (Strukturgenomprojekt)
- European Molecular Biology Laboratory (EMBL), Hamburg
und Heidelberg, Germany
- Max-Planck-Gesellschaft zur Förderung der Wissenschaften
e. V., München, Germany
- Max-Planck-Institut für Infektionsbiologie, Berlin, Germany
- Technische Universität München, Wissenschaftszentrum
Weihenstefan, Freising, Germany
- Combinature Biopharm AG, Berlin, Germany
- Marresearch GmbH, Norderstedt, Germany
- Biomax Informatics AG, Martinsried, Germany
Forschungs- und Entwicklungsvereinbarung
- IKOSATEC GmbH, Garching, Germany
- Combinature Biopharm AG, Berlin-Buch, Germany
Kooperationsvereinbarung: Strukturanalyse mit hohem
Druck für medizinisch relevante Proteine
- Proteinstrukturfabrik: Projekt des Bundesministeriums
für Bildung und Forschung, Germany
Kooperationsvereinbarung zur Erbringung von Verwertungsleistungen
- Ascenion GmbH, München, Germany
Kooperationsvereinbarung über die Fortsetzung der
Öffentlichkeitsarbeit auf dem Forschungscampus Berlin-
Buch
- BBB Management GmbH, Berlin, Germany
Kooperationsvereinbarung: (Forschungs- und Entwicklungsvereinbarung)
Identifizierung von 2-D-separierten
Proteinen über In-Gel-Verdau, Massenspektrometrie und
Datenbankanalyse
- Bundesinstitut für Risikobewertung, Berlin, Germany
- Bundesinstitut für Verbraucherschutz und Veterinärmedizin,
Germany
Kooperationsvereinbarung: Massenspektrometrische
Charakterisierung von Fluoreszenz-markierten Glyko- und
Phosphopeptiden
- Biosyntan GmbH, Berlin-Buch, Germany
Forschungs- und Entwicklungsvereinbarung: Development
of isotope labelled media for protein expression in
bacteria, yeast, insect cells and higher organisms
- Cambridge Isotope Laboratories, Massachusetts, USA
Nutzungs- und Dienstleistungsvereinbarung: Gemeinsame
Nutzung von Forschungsgeräten und technologischen
Einrichtungen
- Charité-Universitätsmedizin Berlin, Germany

Kooperationsvereinbarung zur Nutzung von Übertragungsstrecken
als Zugangsleitungen zum Gigabit-Wissenschaftsnetz
G.WiN
- Verein zur Förderung eines Deutschen Forschungsnetzes
e. V., Berlin, Germany
Kooperationsvereinbarung über die Zusammenarbeit im
Rahmen von Projekten
- Freie Universität Berlin, Germany
Kooperationsvereinbarung: Transfer von Naturstoffen,
Derivaten und Analoga
- Hans-Knöll-Institut für Naturstoff-Forschung e. V. , Germany
Collaboration Agreement mit der AG Oschkinat
- Max-Delbrück-Centrum für Molekulare Medizin, Berlin
Kooperationsvereinbarung: Molecular Fingerprinting of
the Blood-Brain Barrier in Hypoxia-Targeting Brain Vessels
to Treat Stroke
- National Research Council of Canada, Montreal
Infrastrukturnutzungsvereinbarung
- PFS biotech AG, Berlin, Germany
Kooperationsvereinbarung: Nutzung der Primärdatenbank
PD Dr. IE Blasig
- Ressourcenzentrum für das Deutsche Humangenomprojekt,
Germany
Kooperationsvereinbarung für Auftragsmessungen
Prof. Dr. H. Oschkinat
- Schering AG, Berlin, Germany
Kooperationsvereinbarung: Structure determination by
NMR
- University of Oxford, UK
Kooperationsvereinbarung
- The University Wisconsin, Madison, USA
Infrastrukturnutzungsvereinbarung
Combinature Biopharm AG, Berlin, Germany
Kooperationsvereinbarung China-Konsortium
- Deutsches Diabetes Zentrum (DDZ) an der Heinrich-
Heine-Universität, Düsseldorf, Germany
- Deutsches Primatenzentrum GmbH (DPZ), Göttingen
- Forschungszentrum Borstel (FZB) – Zentrum für Medizin
und Biowissenschaften, Borstel, Germany
- Hans-Knöll-Institut für Naturstoff-Forschung (HKI),
Jena, Germany
- Heinrich-Pette-Institut für Experimentelle Virologie und
Immunologie (HPI) , Germany
- Leibniz-Institut für Neurobiologie IfN, Magdeburg, Germany
- Leibniz-Institut für Molekulare Biotechnologie (IMB),
Jena, Germany
- Institut für Pflanzenbiochemie (IPB), Halle a.d. Saale,
Germany
- Institut für Pflanzengenetik und Kulturpflanzenforschung
(IPK), Gatersleben, Germany
Appendix
133

MEETINGS, WORKSHOPS, SYMPOSIA
2003/2004
WISSENSCHAFTLICHE VERANSTALTUNGEN
2003
6th Symposium Signal Transduction in the Blood-Brain
Barriers
Blasig IE, Haseloff RF
Szeged/Ungarn
September 2003
Berlin Magnetic Resonance Seminar and High-Field NMR
Facility Users Meeting
Oschkinat H
Forschungsinstitut für Molekulare Pharmakologie, Berlin-
Buch, Germany
December 2003
4th Progress report meeting, EU-Project: Anti-diuresis
using short-acting vasopressin-V2-receptor agonists as a
new therapeutic strategy of urinary incontinence and voiding
disorders
Klussmann E, Rosenthal W, Lauterjung U
Magnus-Haus, Berlin, Germany
June 2003
1st Progress report meeting, EU-Project: Anchored cAMP
signalling / RTD grant - Implication for treatment of human
diseases
Klussmann E, Rosenthal W, Lauterjung U
Magnus-Haus, Berlin, Germany
June 2003
6. Deutsches Peptidsymposium (mit internationaler Beteiligung)
Bienert M, Dreissigacker M, Dathe M
Humboldt-Universität zu Berlin, Germany
March 2003
8th International Dahlem Symposium on Cellular Signal
Recognition and Transduction, Berlin
Rosenthal W
Charite - Universitätsmedizin Berlin, Germany
June 2003
2004
34. Jahrestagung der Deutschen Gesellschaft für Immunologie
Horak I
Berlin, Henry Ford Building, Germany
September 2004
Festsymposium zur Einweihung des 900-MHz-Spektrometers
am FMP
Oschkinat H, Steuer A, Maul B
Max-Delbrück-Communications Center, Berlin-Buch, Germany
July 2004
7th International Symposium on Signal Transduction in the
Blood-Brain Barriers
Blasig IE, Haseloff RF
Potsdam, Germany
September 2004
EMBO Practical Course "Multidimensional NMR in Structural
Biology II”
Oschkinat H (Co-Organisator)
Ciocco, Italy
September 2004
25. Tagung des Max-Bergmann-Kreises
Bienert M
Munster, France
October 2004

WORK IN PANELS 2003/2004
GREMIENARBEIT 2003/2004
Walter Rosenthal
- Mitglied im Kuratorium des Deutschen Instituts für
Ernährungsforschung, Potsdam-Rehbrücke
- Stellvertretender Sprecher der Sektion C Lebenswissenschaften
der Leibniz Gemeinschaft
- Mitglied des Wissenschaftlichen Beirats des Hans-
Knöll-Insituts, Jena
- Kuratoriumsmitglied der Berlin-Brandenburgischen
Fortbildungsakademie (BBFA)
- Leiter des Instituts für Pharmakologie, Charité-Universitätsmedizin
Berlin
- Schatzmeister im Verbund biowissenschaftlicher und
biomedizinischer Gesellschaften (vbbm)
Hartmut Oschkinat
- Mitglied des Aufsichtsrats der Firma PSF Bio AG, Berlin
Ivan Horak
- Wissenschaftlicher Direktor der Forschungseinrichtung
für Experimentelle Medizin, FU Berlin
- Ombudsmann für Gute Wissenschaftliche Praxis des
Forschungsverbundes Berlin e.V.
- Sprecher der Arbeitsgruppe Tierhaltung der Gemeinsamen
Kommission nach § 3 (3) UniMedG
- Mitglied des Beirats in der Transgenic Core Facility der
RCC Gen bio tec GmbH, Campus Berlin-Buch
Michael Bienert
- Wissenschaftlicher Sekretär des Max-Bergmann-Kreises
REVIEW ACTIVITIES 2003/2004
GUTACHTERTÄTIGKEIT 2003/2004
Bienert, Michael
Organisations
- Deutsche Forschungsgemeinschaft
Research Institutions
- Universität Halle
- Humboldt-Universität zu Berlin
Journals
- Org Lett
- Drug Design Review-Online
- Tetrahedron Lett
- J Mass Spec
- Amino Acids
- J Pept Sci
- J Am Chem Soc
- J Mol Recognit
- Pept Res
Blasig, Ingolf
Organisations
- Deutsche Forschungsgemeinschaft
- Research Grant Council Hong Kong
- Jubiläumsfond Österreichische Nationalbank
- Assoziazone Italiana per la Ricerca Cancro
Research Institutions
- Philipps-Universität Magdeburg
- Universität Potsdam
- Charité-Universitätsmedizin Berlin
Journals
- Neurochemistry
- Glia
- J Neurosci
- J Pharm Pharmacol
- J Neurochem
- Neurochemical Research
Carstanjen, Dirk
Journals
- Bone marrow transplantation
Dathe, Margitta
Organizations
- The Israel Science Foundation
Journals
- J Pep Res
- Eur J Biochem
- Biochim Biophysica Acta
Appendix
135

- Comb Chem
- Biophys J
- J Biol Chem
- New J Chem
- Biochem J
- Regul Peptides
- Org Biomol Chem
Freund, Christian
Organisations
- University of Oxford
Journals
- Regul Peptides
Hagen, Volker
Research Institutions
- Bayrische-Maximilians Universität Würzburg
Journals
- Angew Chemie
- ChemBioChem
- J Am Chem Soc
- Blood
- Eur J Med Chem
- J Mol Struct
Haseloff, Rainer
Journals
- J Photochemistry and Photobiology B-Biology
- Free Radicals Research
Horak, Ivan
Organisations
- Deutsche Forschungsgemeinschaft
- Deutsche Krebshilfe
- Boehringer Ingelheim Fonds
Research Institutions
- Yorkshire Cancer Research, UK
- Universität zu Lübeck
- Taussig Cancer Center, The Cleveland Clinic Foundation
Journals
- J Biol Chem
- Immunity
- Eur J Immunol
- J Mol Cell Biol
- Blood
- Nucleic Acids Research
- Journal Leukemia
Klussmann, Enno
Organisations
- Health Research Board, Ireland
Journals
- JCI
- Glia
- Biol of the Cell
- Biol Cell
- Neuroscience Letters
- J Cell Sci
- Kidney Int
- J Biol Chem
- Eur J Cell Biol
Krause, Eberhard
Journals
- J Anal Chem
- J Pept Sci
- J Mass Spec
- Biochemistry
Krause, Gerd
Journals
- QSAR Journal
- J Comput Aid Mol Des
Meyer, Thomas
Research Institutions
- Georg-August-Universität Göttingen
Journals
- Circulation
Oehlke, Johannes
Journals
- Biochim Biophys Acta
- Eur J Biochem
Oschkinat, Hartmut
Organisations
- Deutsche Forschungsgemeinschaft
- Österreichische Akademie der Wissenschaften
- Schering AG
- Swiss Federal Institute of Technology Zürich
- Studienstiftung Bonn
- Schweizerischer Nationalfond zur Förderung von
Wissenschaftlicher Forschung
- FWF Der Wissenschaftsfond

Research Institutions
- Freie Universität Berlin
- Ludwig-Maximilians-Universität München
- Bayerische-Maximilians Universität Würzburg
- Humboldt-Universität zu Berlin
- Ecole Normale Supérieure de Lyon
- Charité-Universitätsmedizin Berlin
- Eidgenössische Technische Hochschule Zürich
Journals
- J Mol Biol
- J Magn Reson
- J Biol NMR
- J Am Chem Soc
- Nat Struct Biol
- Science
- Biochemistry
- Proc Natl Acad Sci U.S.A.
Piontek, Jörg
Journals
- GLIA
Pohl, Peter
Organisations
- Wellcome Trust
Research Institutions
- Christian-Albrechts-Universität zu Kiel
- Humboldt-Universität zu Berlin
Journals
- Biochemistry
- Biophys J
- Eur J Biochem
- J Bioelectrochemical Society
- BioMed Central
- TIPS
- Proc Natl Acad Sci U.S.A.
Richter, Regina
Journals
- Vasc Pharmacol
- Eur J Pharmacol
- J Cardiovasc Pharma
- Psychopharmacology
Rosenthal, Walter
Organisations
- Deutsche Forschungsgemeinschaft
- Ärztekammer Berlin
- Fakulte de Science Chile
- Boehringer Ingelheim
- Österreichischer Wissenschaftsfond
- Alexander von Humboldt Stiftung
- Ministerio Dell`Istruzione, Dell ùniversita` E Della Ricera
Research Institutions
- Freie Universität Berlin
- Technische Universität Berlin/DRFZ
- Philipps-Universität Marburg
- Eidgenössische Technische Hochschule Zürich
- Eberhard-Karls-Universität Tübingen
- Institut für Zoo- und Wildtierforschung
- Université Montpellier
- Bayerische Maximilians-Universität München
- Christian-Albrechts-Universität zu Kiel
- Universität des Saarlandes
- Oregon Health and Science University Portland
- Fachhochschule Lausitz
Journals
- Neuroscience Letters
- Nephrology Dialysis Transplantation
- Human Molecular Genetics
- Biology of the cell
- Pharmacol & Toxicol
- Hormone Research
- Encyclopedia of Biological Chemistry
- Eur J of Cell Biol
- EJB
- Eur J Biochem
- Eur Biophys J Biophys
- FEBS Lett
- The Lancet
- Kidney Int
Schmieder, Peter
Organizations
- Studienstiftung des deutschen Volkes
Journals
- ChemBioChem
- J Magn Reson
- Chemistry (Wiley VCH)
Schülein, Ralf
Journals
- FEBS Lett
- Biotechniques
Appendix
137

Siems, Wolf-Eberhard
Journals
- J Mol Med
Utebergenov, Darkhan
Journals
- J Neurochem
Vinkemeier, Uwe
Organizations
- European Molecular Biology Organizations
- Boehringer Ingelheim Fonds
- Österreichische Akademie der Wissenschaften
- DAAD
Journals
- Biochim Biophys Acta
- Biochem Biophys Res Comm
- Dev Cell
- EMBO J
- Eur J Biochem
- FEBS Left
- J Cell Biol
- Mol Cell Biol
- Nucleic Acids Res
- Proc Natl Acad Sci U.S.A.
ACADEMIC TEACHING 2003/2004
LEHRE 2003/2004
Beyermann, Michael
Biophysik-Praktikum zum Biacore-Gerät
Freie Universität Berlin
Bienert, Michael
Vorlesung Proteine und Peptide
Humboldt-Universität zu Berlin
Vorlesung zum Biophysikpraktikum (Biacore-Gerät)
Freie Universität Berlin
Blasig, Ingolf
Vorlesung Funktionelle Biochemie
Universität Potsdam
Mastercourse „Medical Neurosciense“. Reguläre Vorlesung,
Seminare und Praktikum
Universität Potsdam
Mastercourse „Medical Neurosciense“. Fakultative Vorlesung
und Seminar
Universität Potsdam
Dathe, Margitta
Biophysik für Studenten der Biochemie: CD-Spektroskopie
Freie Universität Berlin
Freund, Christian
Vorlesung Protein Engineering
Freie Universität Berlin
Vorlesung Molekulare Immunologie
Freie Universität Berlin
Horak, Ivan
Vorlesung Knock-Out-Mäuse
Freie Universität Berlin
Keller, Sandro
Vorlesung Biophysik für Studenten der Biochemie: Isotherm
Titration Calorimetry
Freie Universität Berlin
Klussmann, Enno
Kursus der Allgemeinen Pharmakologie und Toxikologie
Charite-Universitätsmedizin Berlin
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion:
A-Kinase-Ankerproteine
Charite-Universitätsmedizin Berlin

Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion:
Affinitätspräzipationstechniken
Charite-Universitätsmedizin Berlin
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
für Biochemiker, Biologen, Mediziner und
Pharmazeuten
Charite-Universitätsmedizin Berlin
Kursus der Allgemeinen Pharmakologie und Toxikologie
für Humanmediziner
Charite-Universitätsmedizin Berlin
Krause, Eberhard
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
– Massenspektrometrie Proteinanalytik (Proteomics)
Charite-Universitätsmedizin Berlin
Krause, Gerd
Vorlesung Grundlagen Molecular Modelling
Technical University of Applied Sciences, Berlin
Vorlesung Grundlagen Molecular Modelling
Technical University of Applied Sciences, Berlin, FB Bioinformatics
Krause, W
Spezialkurs Sucht
Charite-Universitätsmedizin Berlin
Psychopharmakologie im Praktikum Molekulare und
zelluläre Signaltransduktion
Charite-Universitätsmedizin Berlin
Lorenz, Dorothea
Kursus Mikroskopische Techniken (Intrazelluläre Ca2+-
Messungen)
Humboldt-Universität zu Berlin
Meyer, Thomas
Vorlesung Innere Medizin für Zahmediziner
Universität Göttingen
Praktikum Innere Medizin
Universität Göttingen
Vorlesung Pathologie und Klinik internistischer und chirurgischer
Erkrankungen
Universität Göttingen
Oschkinat, Hartmut
Vorlesung Biologische NMR-Spektroskopie
Freie Universität Berlin
Vorlesung Biophysikalische Methoden
Freie Universität Berlin
Vorlesung Grundlagen und neue Techniken der
biologischen NMR-Spektroskopie
Freie Universität Berlin
Vorlesung Grundlagen der biologischen NMR-Spektroskopie
Freie Universität Berlin
Pohl, Peter
Vorlesung Mikroskopie
Humboldt-Universität zu Berlin
Vorlesung Experimentelle Biophysik
Humboldt-Universität zu Berlin
Vorlesung Biomechanik
Humboldt-Universität zu Berlin
Vorlesung Biophysik III
Johannes-Kepler Universität Linz
Vorlesung Biophysik I
Johannes-Kepler Universität Linz
Übungen zur Physik
Johannes-Kepler Universität Linz
Rademann, Jörg
Vorlesung Heterocyclen
Universität Tübingen
Vorlesung Stereochemistry
Freie Universität Berlin
Vorlesung Combinatorial Chemistry
Quaid-i-Azzam University, Islamabad
Vorlesung Chemie für Biologen
Universität Tübingen
Vorlesung Aktuelle Methoden der Bioorganischen
Synthese
Freie Universität Berlin
Vorlesung Bioorganic and Natural Product Chemistry
Freie Universität Berlin
Richter Regina
Vorlesung Anwendung der Mikrodialysetechnik in der
Neuropharmakologie
Charite-Universitätsmedizin Berlin
Appendix
139

Rosenthal, Walter
Vorlesung Gentherapie I
Charite-Universitätsmedizin Berlin
Vorlesung Weitere alternative Therapieformen
Charite-Universitätsmedizin
Vorlesung Pharmakokinetik
Charite-Universitätsmedizin Berlin
Vorlesung Arzneimittel-Metabolismus/Pharmakogenetik
Charite-Universitätsmedizin Berlin
Vorlesung Gentherapie II (Mechanismen)
Charite-Universitätsmedizin Berlin
Vorlesung Stammzellen
Charite-Universitätsmedizin Berlin
Vorlesung Homöopathie
Charite-Universitätsmedizin Berlin
Vorlesung Placebo
Charite-Universitätsmedizin Berlin
Vorlesung Antivirale Therapie, Antigenese, Gentherapie
Charite-Unviersitätsmedizin Berlin
Schmieder, Peter
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
Freie Universität Berlin
Vorlesung Multidimensionale NMR-Spektroskopie-Grundlagen
und Anwendungen in der Strukturaufklärung
Technische Universität Berlin
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
Technische Universität Berlin
Grundlagen und Anwendungen der Mehrdimensionalen
NMR-Spektroskopie
Technische Universität Berlin
Schülein, Ralf
Kursus der Allgemeinen Pharmakologie und Toxikologie
Charite-Universitätsmedizin Berlin
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
für Biochemiker, Biologen, Mediziner und
Pharmazeuten
Chartie-Universitätsmedizin Berlin
Kursus der allgemeinen Pharmakologie
Charité-Universitätsmedizin Berlin
Vorlesung Molekulare Pharmakologie und zelluläre Signaltransduktion
Charité-Universitätsmedizin Berlin
Vinkemeier, Uwe
Vorlesung Aktuelle Themen der zellulären Siganltransduktion
Freie Universität Berlin
Vorlesung Mechanismen der molekularen Signalverarbeitung
Freie Universität Berlin
Praktikum Mechanismen der Signalverarbeitung
Freie Universität Berlin
Wiesner, Burkhard
Laser Scanning Mikroskopie: Möglichkeiten und Grenzen
optischer Methoden zur Untersuchung subzellulärer Prozesse
Freie Universität Berlin
Konfokale Mikroskopie (Molekulare Pharmakologie und
zelluläre Signaltransduktion)
FU/FMP
Molekulare Pharmakologie und zelluläre Signaltransduktion
FU/FMP
Mikroskopische Techniken (FRAP, FREt, Reflexionsmessungen)
HU/FMP
Laser Scanning Mikroskopie: Möglichkeiten und Grenzen
optischer Methoden zur Untersuchung subzellulärer Prozesse
Freie Universität Berlin

CALLS FOR APPOINTMENTS 2003/2004
RUFE 2003/2004
Hermosilla, Ricardo (2003)
Juniorprofessur für Pathologie der Signaltransduktion
am Institut für Pharmakologie der Charite-Universitätsmedizin
Berlin, Germany
Pohl, Peter (2004)
Lehrstuhl für Biophysik
Technisch-Naturwissenschaftliche Fakultät der Johannes-
Kepler-Universität Linz
Pires, Ricardo (2002)
Associated Professor for Biochemistry and Structural Biology
Instituto De Ciencas Biomédicas, Universidade Federal do
Rio de Janeiro, Brasil
POSTDOCTORAL LECTURE QUALIFICATIONS
2003/2004
HABILITATIONEN 2003/2004
Oksche, Alexander (2003) Molekulare Grundlagen angeborener
und erworbener polyurischer Störungen
Charité-Universitätsmedizin Berlin
Schülein, Ralf (2003) The early secretory pathway of membrane
proteins: clinical and pharmacological implications
Charité-Universitätsmedizin Berlin
141 Appendix

GRADUATIONS 2003/2004
PROMOTIONEN 2003/2004
Alken, Martina (2004) Functional significance of N-terminal
signal peptides of G protein-coupled receptors
Freie Universität Berlin
Andreeva, Anna (2004) Protein kinase C isoform antagonism
controls occludin phosphorylisation and tight
junction assembly
Freie Universität Berlin
Barth, Michael (2004) Entwicklung neuer, hochbeladener
Trägermaterialien für die organische Festphasensynthese
auf Basis von vernetztem Polyethylenimin – Anwendung im
Bereich der Peptidsynthese, für Polymerreagenzien und
zur Synthese von peptidfunktionalisierten Dendrimeren
Eberhard Karls Universität Tübingen
Begitt, Andreas (2004) Nukleocytoplasmatischer Transport
und Geninduktion durch den Transkriptionsfaktor STAT1
Freie Universität Berlin
Boisguerin, Prisca (2004) Characterization of PDZ Domain/
Ligand Specifity
Freie Universität Berlin
Castellani, Federica (2004) Structure determination of
immobilized proteins by solid-state NMR spectroscopy
Freie Universität Berlin
Eckhardt, Torsten (2004) Design, Synthese, Photochemie
und biologische Anwendung von ausgewählten „caged“
cyclischen Adenosin-3, 5´-monophosphaten
Humboldt-Universität zu Berlin
Edemir, Bayram (2004) Klonierung und Charakterisierung
einer neuen AKAP18-Isoform, AKAP18δ, und ihre mögliche
Beteiligung an der AVP-vermittelten AQP2-Translokation
Freie Universität Berlin
Meyer, Thomas (2004) Nukleäre Akkumulation und Zielgenerkennung
von STAT1-Transkriptionsfaktoren
Freie Universität Berlin
Osiak, Anna (2004) Die Rolle der ubiquitinähnlichen Gene
UBL 14 und ISG15 in vivo
Freie Universität Berlin
Sauer, Ines (2004) Apolopoprotein E - abgeleitete Peptide
als Vektoren zur Überwindung der Blut-Hirn-Schranke
Freie Universität Berlin
Serowy, Steffen (2004) Migration von Protonen entlang der
Oberfläche ebener Bilipidmembranen
Humboldt-Universität zu Berlin
Thielen, Anja (2004) Identifizierung transportrelevanter
Aminosäurereste im proximalen C-Terminus des humanen
Vasopressin-V2-Rezeptors
Freie Universität Berlin
Storm Robert (2004) Extracellular osmolality and solute
composition participate in the expressional regulation of
Aquaporin-2 in renal inner medullary collecting duct cells.
Evidence for an involvement of the TonE/TonEBP pathway
Freie Universität Berlin
Wessolowski, Axel (2004) Amphipathische Hexapeptide -
Interaktion mit Membranen
Freie Universität Berlin
Zimmermann, Jürgen (2004) Struktur und Funktion von
EVH1-Domänen
Freie Universität Berlin
Zühlke, Kerstin (2003) Strukturelle und funktionelle Bedeutung
der konservativen Disulfidbrücke des Vasopressin-
V2-Rezeptors
Freie Universität Berlin

DIPLOMA THESES 2003/2004
DIPLOMARBEITEN 2003/2004
2003
Brandenburg, Martin
Kerntransportverhalten trunkierter STAT-Porteine
Technische Fachhochschule Berlin
Hahn, Janina
Untersuchungen zu Chromophor-Protein-Interaktionen in
ortsspezifischen Mutanten von Phytochrom Cph1 aus Cyanobakterium
Synechocystis
Freie Universität Berlin
Heine, Markus
Wasserstoffperoxyd-induzierte Veränderungen der Aktivität
und Expression ausgewählter Proteine in Epithelzellen
Freie Universität Berlin
Paschke, Carmen
Erweiterte Peakformanalyse zur Verbesserung der automatischen
Resonanzzuordnung von NMR-Spektren
Fachhochschule Lausitz
Schröder, Stephan
Derivatisierung phosphorylierter Peptide zur Erhöhung der
Signalintensität in der Massenspektrometrie
Technische Universität Berlin
Thurisch, Boris
Strategien zur gezielten Expressions-Inhibition des murinen
FMIP-Gens
Technische Fachhochschule Berlin
Wendt, Norbert
Strukturuntersuchungen der Disulfidstruktur an N-terminalen
CRF-Rezeptoren
Technische Universität Berlin
2004
Cubic, Ivona
Charakterisierung von Proteinen mittels Kapillar-HPLC und
Massenspektrometrie
Technische Fachhochschule Berlin
El-Dashan, Adeeb
Milde C-Acylierungen von polymergebundenen Carboxylatphosphoranen
für Anwendungen in der Medizinischen
Chemie
Eberhard-Karls-Universität Tübingen
Kotzur, Nico
Synthese und Photochemie von photolabilen cAMP-Derivaten
Humboldt-Universität zu Berlin
Kubald, Sybille
Extrazelluläre Domäne des Rezeptors für den Corticotropin-Releasing-Faktor:
Klonierung in Expression eines
N-Terminus-Schleife-Konstrukts als Bindungsmodell
Technische Fachhochschule Berlin
Lassowski, Brigitte
Untersuchungen zur Clonierung und Reinigung von Tight-
Junction-Proteinen
Universität Potsdam
Renner, Armin
Funktionelle Bedeutung des Signalpeptides des Corticotropin-Releasing-Faktor
Rezeptors 2a
Julius-Maximilians-Universität Würzburg
Roswadowski, Inga
Untersuchungen zur Struktur und Funktion von Tight-
Junction-Proteinen
Technische Fachhochschule Berlin
Schröder, Stephan
Derivatisierung phophorylierter Peptide zur Erhöhung der
Ionisierungen der Massenspektrometrie
Technische Universität Berlin
Zech,Tobias
Functional studies of the adapter protein CD2BP2
Freie Universität Berlin
143 Appendix

INTERNSHIPS 2003/2004
PRAKTIKANTEN 2003/2004
Ballmer, Boris
10.11.2003–05.12.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Berger
Behling, Katja
01.06.2004–25.06.2004
Freie Universität Berlin – Biochemie
Supervision: Dr.Berger
Behnken, Swantje
16.08.2004–08.10.2004
Universität Potsdam – Biochemie
Supervision : Dr. Donalies
Bibow, Stefan
01.05.2004–31.07.2004
Humboldt-Universität zu Berlin – Biophysik
Supervision: Prof. Reif
Bieberstein, Andrea
06.10.2003–20.02.2004
Fachhochschule Lausitz – Biotechnologie
Supervision: Dr. Donalies
Blick, Kristin
12.05.2003–20.06.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Berger
Böthe, Matthias
08.12.2003-30.01.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Böthe, Matthias
01.07.2004–11.08.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Dathe
Brückner, Kathrin
07.04.2003–09.05.2003
Technische Universität Dresden – Chemie
Supervision: Dr. Siems
Christian, Frank
15.09.2003–24.10.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Klussmann
Cubic, Ivona
01.09.2003–29.02.2004
Technische Fachhochschule – Biochemie
Supervision: Dr. E. Krause
Curth, Sebastian
14.06.2004–10.09.2004
Technische Fachhochschule Berlin – Biotechnologie
Supervision: Dr. Donalies
Elß, Franka
04.10.2004–25.03.2005
Fachhochschule Magdeburg – Pharmakologie
Supervision: Dr. E. Krause
Erel, Funda
01.02.2003–31.03.2003
Vorpraktikum Biotechnologie
Supervision: Dr. Klußmann
Ernst, Oliver
20.10.2003–19.04.2004
Technische Universität Berlin – Biotechnologie
Supervision: Dr. Donalies
Femmer, Christian
01.08.2004–15.10.2004
Technische Universität Berlin – Biotechnologie
Supervision: Dr. Klose
Gartmann, Marco
20.10.2003–31.12.2003
Universität Potsdam – Biochemie
Supervision: Dr. Haseloff
Gasser, Carlos
14.05.2004–16.07.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Pohl
Giebel, Sebastian
01.06.2004-16.07.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Groß, Anika
02.02.2004–12.03.2004
Freie Universität Belin – Pharmazie
Supervision: Dr. Schmieder
Güngör, Volkan
01.02.2004–30.04.2004
Berufsanfänger
Supervision: H.-J. Mevert

Guse, Katrin
19.04.2004-04.06.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Han, Soeng-Ji
19.07.2004-03.09.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Hamidzadek, Nadja
27.01.2003–27.03.2003
Weiterbildung (Arbeitsamt) – Biochemie
Supervision: Dr. Labudde
Heinze, Mathias
01.02.2004–31.03.20004
Humboldt-Universität zu Berlin
Supervision: T. Jahn
Heuberger, Julian
08.11.2004–14.12.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Dathe
Hübner, Florian
01.11.2004-18.02.2005
Technische Fachhochschule Berlin
Supervision: Dr. Pohl
Hühn, Stephan
29.03.2004-07.05.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Jahnke, Nadine
01.06.2004-16.07.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Jehle, Stefan
11.08.2003–24.10.2003
Freie Universität Berlin – Biochemie
Supervision: Prof. Oschkinat
Kalmbach, Norman
06.01.2003–31.01.2003
Technische Fachhochschule Berlin – Biotechnologie
Supervision: Dr. Vinkemeier
Kaltofen, Sabine
21.07.2003–10.10.2003
Fachhochschule Zittau
Supervision: Dr. Krabben
Kamitz, Anne
08.01.2004–02.04.2004
Abiturientin
Supervision: Dr. Vinkemeier
Kieseritzky, Gernot
01.03.2004–09.04.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Pohl
Kirsch, Jenny
13.04.2004–30.04.2005
Universität Potsdam – Biochem.
Supervision: Dr. Blasig
Kotzur, Nico
01.03.2004–31.12.2004
Humboldt-Universität Berlin – Chemie
Supervision: Dr. Hagen
Kraemer, Florian
01.08.2004–24.09.2004
Universität Potsdam – Biochemie
Supervision: Prof. Horak
Kron, Anja
01.09.2003–30.01.2004
Fachhochschule – Biotechnologie
Supervision: Dr. Leitner
Kronsbein, Helena
10.03.2003–28.03.2003
Technische Universität München – Biochemie
Supervision: Dr. Schmieder
Krüger, Kerstin
15.04.2003–14.10.2003
Rharmazie-Praktikantin
Supervision: Dr. Siems
Krüger, Magnus
01.11.2004–30.04.2005
Freie Universität Berlin – Pharmazie
Supervision: Dr. Beyermann
Kuhn, Ramona
04.08.2003–26.09.2003
Technische Universität – Berlin – Umweltschutz
Supervision: Dr. Wiesner
Lassowski, Birgit
01.01.2003–31.03.2003
Universität Potsdam – Biotechnologie
Supervision: Dr. Blasig
145 Appendix

Leddin, Mathias
01.08.2003–31.01.2004
Freie Universität Berlin – Bio
Supervision: Dr. Knobeloch
Lisewski, Ulrike
14.04.2004–18.05.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Pohl
Marter, Katrin
05.05.2004–31.10.2004
Freie Universität Berlin – Biologie
Supervision: Dr. Haseloff
Mehmedi, Burhan
24.11.2003–26.12.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Siems
Mitschke, Doreen
02.06.2003–11.07.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Klußmann
Müller, Jeanette
19.04.2004-04.06.2004
Freie Universität Berlin – Biochemie
Supervision: S. Keller
Narayanan, Raghav
17.05.2004–07.08.2004
Neu Delhi
Supervision: Dr. Freund
Neumann, Christine
23.02.2004–02.04.2004
Universität Marburg – Biologie
Supervision: Dr. Vinkemeier
Neumann, Marleen
03.02.2003–08.03.2003
Technische Fachhochschule Berlin – Biotechnologie
Supervision: Dr. Wiesner
Niehage, Christian
01.08.2004–31.10.2004
Freie Univerisät Berlin – Biochemie
Supervision: Dr. Blasig
Oehlke, Elisabeth
12.07.2004–30.08.2004
Freie Universität Berlin – Chemie
Supervision: Dr. Hagen
Overath, Thorsten
01.09.2003–26.10.2003
Technische Universität Berlin – Biotechnologie
Supervision: Dr. Siems
Overath, Thorsten
23.02.2004–14.05.2004
Technische Universität – Biotechnologie
Supervision: Dr. Siems
Petrovska, Silvija
15.11.2004–10.12.2004
Skopje-Mazedonien
Supervision: Dr. Siems
Pietske, Matthias
19.07.2004-03.09.2004
Universität Potsdam – Biochemie
Supervision: S. Keller
Prigge, Matthias
24.09.2003–07.11.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Pohl
Reibitz, Franziska
01.02.2004–31.07.2004
Pharmazeutin
Supervision: Dr. Siems
Roswadowski, Inga
01.06.2004–30.09.2004
Technische Fachhochschule Berlin – Biotech.
Supervision: Dr. Blasig
Santamaria, Katja
19.02.2003–28.02.2003
Freie Universität Berlin
Supervision: Dr. Klussmann
Schäfer, Zasia
06.09.2004–08.10.2004
Universität Potsdam – Biochemie
Supervision: Dr. Carstanjen
Schlede, Stephanie
04.08.2003–29.08.2003
Universität Potsdam – Biotechnologie
Supervision: Dr. Siems
Schmidt, Marco
08.12.2003–31.01.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Hagen

Schröder, Kati
24.02.2003–14.03.2003
Technische Universität Berlin – Biotechnologie
Supervision: M. Alken
Schulz, Katrin
01.12.2004–28.02.2005
Charitè-Universitätsmedizin Berlin
Supervision: Dr. Blasig
Schumacher, Anne
04.08.2003–12.09.2003
Humboldt-Universität zu Berlin – Biologie
Supervision: Dr. Krabben
Schumacher, Anne
01.06.2004–31.07.2004
Freie Universität Berlin – Biologie
Supervision: Dr. Diehl
Schumann, Franziska
15.09.2004–28.02.2005
Technische Fachhochschule Berlin. – Chemietechnologie
Supervision: Dr. Beyermann
Schuster, Ariane
01.07.2004–30.04.2005
Universität Potsdam – Biochemie
Supervision: Dr. Blasig
Seifert, Christoph
03.03.2003–28.03.2003
Freie Universität Berlin – Biochemie
Supervision: Dr. Siems
Socher, Elke
17.02.2003–19.03.2003
Humboldt-Universität zu Berlin – Chemie
Supervision: Dr. Hagen
Stengel, Florian
25.05.2004–06.07.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Siems
Stenzel, Denise
01.09.2003–30.01.2004
Fachhochschule Lausitz – Biotechnologie
Supersision: Dr. Knobeloch
Strohschein, Susan
07.01.2003–15.02.2003
FU Berlin – Biochemie
Supervision: Dr. Blasig
Teodorczyk, Marcin
07.06.2004–27.08.2004
Italien – Biotechnologie
Supervision: Dr. Klussmann
Trippens, Jessica
19.10.2004–11.01.2005
Technische Hochschule Berlin – Biotechnologie
Supervision: Dr. Carstanjen
Vorwinkel, Jakob
08.03.2004–07.05.2004
Freie Universität Berlin – Biochemie
Supervision: Dr. Pohl
Wellmann, Anke
29.03.2004–13.08.2004
Freie Universität Berlin: Biotechnologie
Supervision: Dr. Krabben
Winkler, Franziska
26.07.2004–26.08.2004
Technische Fachhochschule Berlin – Biochemie
Supervision: Dr. Siems
Wolf, Constanze
13.04.2004–12.07.2004
Universität Potsdam – Biologie
Supervision: Dr. Blasig
Wolkenhauer, Jan
01.01.2003–30.06.2003
Universität Potsdam – Biotechnologie
Supervision: Dr. Wiesner
Zuleger, Nikolaj
01.09.2003–30.01.2004
Fachhochschule Lausitz – Biotechnologie
Supervision: Dr. Blasig
Zschörnig, Barbara
25.08.2003–26.09.2003
Universität Regensburg – Biochemie
Supervision: Dr. Vinkemeier
Zimmerling, Katrin
25.10.2004–18.03.2005
Hochschule Anhalt – Pharmazeutische Technologie
Supervision: Dr. Dathe
Appendix
147

GUEST SCIENTISTS 2003/2004
GASTWISSENSCHAFTLER 2003/2004
2003
Antonenko, Juri
30.01.2003–27.02.2003
01.11.2003–30.11.2003
Belozerski Insitut Moscow, Russia
Alexandrov, Alexej
15.03.2003–15.09.2003
01.11.2003–31.05.2004
Kazan University, Russia
Ayuyan, Artem
13.08.2003–30.11.2003
Academy of the Sciences, Moscow, Russia
Balla, Zsolt
01.09.2003–14.09.2003
University of Debrecen, Hungary
Barany-Wallje, Elsa
08.09.2003–29.02.2004
University of Stockholm, Sweden
Lents, Alexander
24.07.2003–28.09.2003
Academy of the Sciences Moscow, Russia
Pechanova, Olga
25.06.2003–20.07.2003
Academy of the Sciences Slovakia, Bratislava
Paulis, Ludovit
17.11.2003–21.11.2003
Academy of the Sciences Slovakia, Bratislava
Pires, Jose R.
01.11.2003–31.01.2004
University of Rio de Janeiro, Brasil
Woineshet, Zenebe
25.06.2003–20.07.2003
Academy of the Sciences Bratislava, Slovakia
Sokolov, Valerij
20.06.2003–18.07.2003
01.10.2003– 31.10.2003
Academy of the Sciences Moscow, Russia
2004
Alexandrov, Alexej
01.11.2003–15.03.2004
Kazan University, Russia
Antonenko, Juri
30.04.2004–30.05.2004
01.10.2004–31.10.2004
Belozerski Institute, University of Moscow, Russia
Andreeva, Anna
01.04.2004–31.05.2004
Russia, Scholarship holder
Arbuzova, Anna
19.04.2004–28.05.2004
University of New York in Stony Brook, USA
Barany-Wallje, Elsa
08.09.2003–29.02.2004
University of Stockholm, Sweden
Coin, Irene
01.10.2004–31.10.2004
Italy, Scholarship holder
Khanfar, Monther
08.07.2004–19.09.2004
University of Zarga, Hashemite, Jordanien Uni
Kumari, Neha
01.05.2004–31.12.2004
India, Scholarship holder
Lents, Alexander
11.08.2004–11.10.2004
Frumkin Institute for Electrochemistry, University of Moscow,
Russia
Magalhaes de Souza, Crista
01.01.2004–30.11.2004
Oswald Criz Institute, Rio de Janeiro, Brasilia
Pires, Richardo
01.11.2003–31.01.2004
01.12.2004–28.02.2005
University of Rio de Janeiro, Brasil
Schreibelt, Gerty
03.05.2004–25.09.2004
VU Medical centre Amsterdam, The Netherlands

Sokolov, Valerij
11.03.2004–11.04.2004
18.10.2004
17.11.2004
Frumkin Institute for Electrochemistry, University of Moscow,
Russia
Sokolenko, Elena
11.08.2004–11.10.2004
Frumkin Institute for Electrochemistry, University of Moscow,
Russia
Sommer, Klaus
15.11.2004–28.01.2005
Universität of Linz, Austria
LECTURES AT THE FMP 2003
KOLLOQUIEN UND SEMINARE AM FMP 2003
Bauer, Hans (Institut für Molekularbiologie, Salzburg,
Austria)
Tight junction proteins
26.02.2003
Host: Blasig IE
Beitz, Eric (Institut für Pharmazeutische Chemie, Universität
Tübingen, Germany)
Aquaporin mediated water permeability in the inner ear
07.10.2003
Host: Klussmann E
Berger, Hartmut (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
GS/GI coupling of the CRF receptors type 1 in HEK cells
14.01.2003
Beyermann, Michael (Forschungsinstitut für Molekulare
Pharmakologie, Berlin-Buch, Germany)
Struktur- und Bindungsstudien extracellulärer Rezeptordomänen
11.03.2003
Blasig IE (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Structure, Function and Regulation of Tight Junction Proteins,
25.11.2003
Blechert, Siegfried (Institut für Chemie der Technischen
Universität Berlin, Germany)
Metathese und Wirkstoffchemie – eine starke Kombination
17.07.2005
Host: Oschkinat H
Carstanjen, Dirk (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Die Rolle des Interferon induzierten Transkriptionsfaktors
ICSBP in der Reifung und Funktion von Zellen des myelopoetischen
Systems
09.12.2003
Danilov, Sergei M (Anestesiology Research Center, Chicago,
USA)
Structure-function studies of angiotensin-converting enzyme
using monoclonal antibodies
24.11.2003
Host: Siems WE
Appendix
149

Dathe, Margitta (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
ApoE-Peptid-Lipidkomplexe: Trägermodelle für eine Wirkstoffaufnahme
ins ZNS
14.10.2003
Dieckmann, Torsten (University of California)
Mechanism of action of Rab GTPases in intracellular vesicular
protein transport
02.07.2003
Host: Oschkinat H
Diehl, Anne (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Protein production for structure determination by NMR
and X-ray
25.02.2003
Duffy, Heather S. (Albert-Einstein College of Medicine,
New York, USA)
pH dependent inter- and intramolecular interactions on
connexin43: Regulation of a junctional complex
02.09.2003
Host: Blasig IE
Folkers, Gerd (Eidgenössische Technische Hochschule
Zürich, Switzerland)
Designing the Locks and Creating new Keys: Molecular
Design Methodology for Genetic Switches
06.05.2003
Host: Bienert M
Freund, Christian (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
GYF and AH3 domain mediated interactions in eukayotic
signaling
10.06.2003
Glockshuber, Rudi (Eidgenössische Technische Hochschule
Zürich, Switzerland)
Catalysis of disulfide bond formation in Escherichia coli
21.01.2003
Host: Bienert M
Goody, Roger (Max-Planck-Institut für Molekulare Physiologie,
Dortmund, Germany)
Mechanisms of actions of Rab GTPases in intracellular
vesicular protein transport
01.07.2003
Host: Oschkinat H
Gottschalk, Kay (Weizmann Institute of Science, Israel)
Computational Approaches to Transmembrane Protein
Structure
27.11.2003
Host: Reif B
Hagen, Volker (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Caged compounds: Design and applications
13.05.2003
Hauri, Hans-Peter (Universität Basel, Switzerland)
Protein traffic early in the secretory pathways: dynamics
and signals
23.09.2003
Host: Hermosilla R
Heinz, Dirk (Department of Structural Biology, Braunschweig,
Germany)
Bacterial invasion at atomic resolution
16.09.2003
Host: Rosenthal W
Heise, Bernd (Institut für Experimentelle Physik der Universität
Ulm, Germany)
Structural investigations on peptaibols via liquid and solid
state NMR
18.03.2003
Host: Oschkinat H
Helm, Volkhard (MPI Frankfurt, Germany)
Studying protein-protein interactions in silico
11.02.2003
Host: Freund C
Hennemann, Hanjo (Center of advanced european studies
and research, Bonn, Germany)
Ras recruitment system: analysis of protein function and
protein networks and its high throughput application
21.10.2003
Host: Rosenthal W
Hermosilla, Ricardo (Forschungsinstitut für Molekulare
Pharmakologie, Berlin-Buch, Germany)
Intracellular degradation pathways of wild-type and transport-defective
mutant vasopressin V2 receptors
09.07.2003
Hougardy, Stefan (Institut für Informatik der Humboldt-Universität
zu Berlin, Germany)
Three-Dimensional similarity of Small Molecules
13.11.2003
Host: Vinkemeier U
Johnson, Nils (Forschungszentrum Karlsruhe, Germany)
Analysis of protein network in living cells
02.12.2003
Host: Vinkemeier U

Jordt, Sven (Department of Cellular & Molecular Pharmacology,
UCSF, USA)
The Pain Gate – Functional Domains in the Capsaicin
Receptor
16.12.2003
Host: Rosenthal W
Klussmann, Enno (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Protein kinase A anchoring proteins and Rho are involved
in the AVP-indused shuttling of aquaporin
11.02.2003
Krabben, Ludwig (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
NMR am Membranprotein
30.09.2003
Krause, Eberhard (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Structural analysis of peptides and proteins by mass spectrometry
22.04.2003
Krause, Gerd (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Predictions of structure-function relationships: Advantages
and limits of structural bioinformatics
08.04.2003
Kühne, Ronald (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Luteinizing Hormone Releasing Hormone Receptor: Molecular
dynamics and model-based ligand design
24.06.2003
Ladizhansky, Vladimir (Francis Bitter Magnet Laboratory,
Cambridge, UK)
Techniques for modern solid-state NMR
10.01.2003
Host: Oschkinat H
Lang, Christine (Institut für Mikrobiologie und Genetik der
Technischen Universität Berlin, Germany)
Production of proteins for structural genomics using yeast
as hosts
30.04.2003
Host: Schade M
Markley, John L. (University of Wisconsin-Madison, USA)
Structural Genomics of the Eukaryote Arabidopsis thaliana
23.09.2003
Host: Oschkinat H
Marx, Dominik (Lehrstuhl für Theoretische Chemie, Ruhr-
Universität Bochum, Germany)
Proton Transfer along Hydrogen-Bonded Networks
20.05.2003
Host: Pohl P
Monsees, Thomas (Zentrum für Dermatologie und Andrologie,
Giessen, Germany)
Expression von Kininrezeptoren im Rattentestis: Eine funktionelle
Bedeutung für die Spermatogenese
13.01.2003
Host: Siems WE
Mourot, Alexandre (Laboratoire De Chimie Bioorganique,
France)
Investigation of the conformational transitions of the nicotinic
receptor by photoaffinity labeling, 12.12.2003
Host: Pohl P
Müller, Jürgen (National Cancer Institute at Frederick,
USA)
C-TAK1 is a regulator of the MAPK scaffold protein KSR
and various other signaling proteins
20.03.2003
Host: Rosenthal W
Multhaup, Gerd (Institut für Chemie und Biochemie, Freie
Universität Berlin, Germany)
Implication of amyloid precursor protein ligands in amyloidogenesis
26.08.2003
Host: Reif B
Palczewski, Krysztof (Department of Ophtalmology of the
University of Washington, USA)
Structure, function and membrane organisation of vertebrate
rhodopsin
10.04.2003
Host: Krause G
Reiser, Oliver (Institut für Organische Chemie der Universität
Regensburg, Germany)
Arglabin, ein vielversprechender Farnesyltransferase-
Inhibitor
13.02.2003
Host: Bienert M
Scharnagl, Hubert (Universitätsklinikum Freiburg, Germany)
Apolipoprotein E, Lipidstoffwechsel und die Alzheimersche
Krankheit
12.06.2005
Host: Dathe M
Appendix
151

Schmalz, Hans-Günther (Universität zu Köln, Germany)
Mediated synthesis of biologycally relevant small molecules
27.03.2003
Host: Oschkinat H
Schmidt, Gundula (Albert-Ludwigs-Universität Freiburg,
Germany)
Mechanism and function of Rho targeting bacterial toxins
19.08.2003
Host: Rosenthal W
Schmieder, Peter (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Strukturen von GPCRs mit Lösungs-NMR
22.07.2003
Schubert, Mario (University of British Columbia in Vancouver,
Canada)
Insights into mRNA degradation in E. coli - The S1 domain
of Rnase E
19.06.2003
Host: Oschkinat H
Schülein, Ralf (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
The early secretory pathway of G protein-coupled receptors:
clinical and pharmacological implications
28.01.2003
Serrano, Luis (European Molecular Biology Laboratory,
Heidelberg, Germany)
Sequence determinants of amyloid formation
20.10.2003
Host: Oschkinat H
Siems, Wolf-Eberhard (Forschungsinstitut für Molekulare
Pharmakologie, Berlin-Buch, Germany)
NEP und ACE, zwei „alte“ Enzyme mit vielen neuen Chancen
16.12.2003
Sinning, Irmgard (Biochemie-Zentrum Heidelberg,
Germany)
Expression of G-protein coupled receptors in the eye of
transgenic Drosophila
04.03.2003
Host: Rosenthal W
Stiles, Brad (Institut für Experimentelle und Klinische
Pharmakologie und Toxikologie, Freiburg, Germany)
Biotoxins: Antrax to Venoms
14.02.2003
Host: Rosenthal W
Stubbs, Milton T. (Institut für Biotechnologie, Martin-
Luther-Universität Halle, Germany)
Understanding protein - ligand interactions: Effects of flexibility
18.02.2003
Host: Oschkinat H
Vinkemeier, Uwe (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Regulation der subzellulären Verteilung von STAT1
27.05.2005
Zamora, Salvador Ventura (Departemente de Bioquimica i
Biologia Molecular, University of Barcelona, Spain)
Short amino acid sequences can trigger protein amyloid
formation in globular proteins
15.07.2003
Host: Reif B

LECTURES AT THE FMP 2004
KOLLOQUIEN UND SEMINARE AM FMP 2004
Ahnert-Hilger, Gudrun (Institut für Anatomie der Charitè-
Universitätsmedizin Berlin, Germany)
Regulation of vesicular neurotransmitter transporters by
heterotrimeric G-proteins
17.02.2004
Host: Klussmann E
Appelt, Christian (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Peptide-lipid interaction: structural investigations by NMR
and Molecular Dynamics
13.04.2004
Baeuerlein, Edmund (Max-Planck-Institut für Biochemie,
Martinsried, Germany)
BIOMINERALISATION. Von Biologie zu Materialwissenschaften
und Molekularbiologie
15.11.2004
Host: Oschkinat H
Banci, Lucia (Centro Risonanze Magnetiche, University of
Florence, Italy)
NMR on Metalloprotein in Structural Genomics
03.05.2004
Host: Oschkinat H
Boelens, Wilbert (University of Njimegen, Department of
Biochemistry, The Netherlands)
Involvement of the small heat shock protein aB-crystallin
in cellular stress regulation
06.04.2004
Host: Reif B
Cordes, Frank (Konrad-Zuse-Zentrum für Informationstechnik,
Berlin, Germany)
Conformation Databases for Virtual Screening
18.05.2004
Host: Kühne R
Gast, Klaus (Max-Delbrück-Center, Berlin-Buch, Germany)
Critical oligomers in protein misfolding and aggregation
10.02.2004
Host: Bienert M
Gershengorn, Marvin (NIH/NIDDK Bethesda, USA)
Pharmacology of Thyrotropin-Releasing Hormone Receptors:
Similarities and Differences
02.03.2004
Host: Krause G
Gohla, Antje (Departments of Immunology and Cell Biology,
La Jolla, USA)
Regulation of cytoskeletal dynamics by Chronophin, a
novel unconventional cofilin phosphatase
30.03.2004
Host: Rosenthal W
Harteneck, Christian (Institut für Pharmakologie der
Charité Universitätsmedizin Berlin, Germany)
Kationenkanäle auf der Suche nach neuen Funktionen: Die
molekulare Vielfalt der TRP-Kanäle
07.09.2004
Host: Rosenthal W
Heise, Tilmann (Heinrich-Pette-Institut Hamburg,
Germany)
The La Protein, a multifunctional RNA binding protein
essential for cell cycle progression
05.10.2004
Host: VinkemeierU
Henning, Mirko (Department of Molecular Biology & The
Skaggs Institute of Chemical Biology, La Jolla, USA)
Labeling Methodology and 19F NMR of Fluorinated RNA
02.06.2004
Host: Reif B
Horuk, Richard (Berlex AG, Richmont, Californien, USA)
CCR1 receptor antagonists from the bench to the Clinic
07.12.2004
Host: Kühne R
Huber, Otmar (Charité Universitätsmedizin Berlin, Germany)
The fate of Cell-cell contacts in apoptotic epithelial cells
15.06.2004
Host: Blasig IE
Hundsrucker, Christian/Weber, Viola (Forschungsinstitut
für Molekulare Pharmakologie, Berlin-Buch, Germany)
Peptide Disruptors of AKAP-PKA Interactions
07.12.2004
John, Susan (King's College, London, UK)
Regulating the transcriptional activity of STAT5
16.11.2004
Host: Vinkemeier U
Kofler, Michael (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
FMP, NWG Protein Engineering
Recognition Code of GYF domains
07.12.2004
Appendix
153

Kungl, Andreas (Universität Graz, Switzerland)
Biophysical Investigations of Chemokine-Receptor and
Co-Receptor Interactions
06.01.2004
Host: Oschkinat H
Labudde, Dirk/Leitner, Dietmar (Forschungsinstitut für
Molekulare Pharmakologie, Berlin-Buch, Germany)
Tools and concepts for automation of protein structure
determination by NMR
09.03.2004
Langer, Gernot (Schering AG, Berlin, Germany)
Early Drug Discovery - Target identification, Assay development
& High throughput screening 16.11.2004
Host: Kühne R
Livett, Bruce G (University of Melbourne, Australia)
Beauty and the Beast: molecular Prospecting for Novel
Drugs from the Sea: the discovery, properties and development
of the novel analgesic, conotoxin Vc1.1 (ACV1)
26.01.2004
Host: Oehme P
Llinas, Miguel (Department of Chemistry of the Carnegie
Mellon University, Pittsburgh, USA)
Is it NMR or is it crystallography? CLOUDS: a direct
method for protein structure elucidation via NMR proton
densities
15.06.2004
Host: Oschkinat H
Lorenz, Dorothea (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Electron microscopy in cell biology. Methods and perspectives
24.02.2004
Luy, Burkhard (Institut für Organische Chemie und Biochemie
II der Technischen Universität München, Germany)
Structural Investigations on the Pulmonary Surfactant
Associated Lipopeptide SP-C and Novel NMR-Techniques
Concerning Small Molecules in Organic Solvents
29.07.2004
Host: Reif B
Marg, Andreas (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
STAT – Abwege
25.05.2004
Mayer, Thomas (Max-Planck-Institute of Biochemistry,
Department of Cell Biology, Martinsried, Germany)
Small molecules: versatile probes to study mitotic kinesins
05.10.2005
Host: von Kries J
Oelgeschläger, Thomas (Marie Curie Research Institut,
Transcription Laboratory, Oxted, UK)
Core promoter-specific regulation of RNA polymerase II
transcription
04.05.2004
Host: Vinkemeier U
Overduin, Michael (University of Birmingham, UK)
Membrane binding domains: the good, the bad, and the
ugly
14.04.2004
Host: Oschkinat H
Pardo, Leonardo (Universidad Autonoma de Barcelona,
Spain)
Bioinformatic Approaches Leading to an Understanding of
the Structure and Activity of G-protein coupled receptors
30.11.2004
Host: Kühne R
Rapp, Wolfgang (Rapp Polymer GmbH, Tübingen,
Germany)
Ein universelles Linkerkonzept zur sequenziellen Abspaltung
von Peptiden und Erzeugung von geschützten Peptidamiden
06.05.2004
Hosts: Bienert M, Beyermann M
Reif, Bernd (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Misfolding proteins and molecular chaperones by solution
and solid-state NMR
10.11.2004
Rosen, Michael (Southwestern Medical Center Dallas,
University of Texas, USA)
Structural and Biochemical Mechanisms of Signal Integration
by the Wiskott-Aldrich Syndrome Protein
25.03.2004
Host: Oschkinat H
Salditt, Tim (Institut für Röntgenphysik, Universität Göttingen,
Germany)
Probing the Structure, and Interactions of Polypeptides in
Lipid Bilayers by X-ray and Neutron Scattering
12.03.2004
Host: Freund C
Scott, John D. (FRS, Oregon Health & Sciences University,
Portland, USA)
The Molecular Architecture of Signal Transduction complexes
05.02.2004
Host: Klussmann E

Seelig, Joachim (Biozentrum Universität Basel, Switzerland)
Detergents, Peptides and Microdomains in Membranes
04.02.2004
Host: Bienert M
Seitz, Oliver (Humboldt-Universität zu Berlin, Germany)
Erzwungene Interkalation – Wie man Basenlücken stopft
und Mutationen in DNA nachweist 19.10.2004
Host: Bienert M
Soderhäll, Arvid (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Suggested function of the antimicrobial cyc-RW peptide
from MD-simulations
22.06.2004
Sommer, Klaus (Medizinische Universität Wien, Austria)
Inhibitor of apoptosis protein survivin is upregulated by
oncogenic c-H-Ras
28.10.2004
Host: Pohl P
Stefan, Eduard (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Phosphodiesterase pde4 is involved in the vasopressinmediated
water reabsorbtion by regulating the localization
of the water channel aquaporin-2
11.05.2004
Strauss, Holger (Forschungsinstitut für Molekulare Pharmakologie,
Berlin-Buch, Germany)
Biophysical investigations on the N-terminus of cyanobacterial
phytochrome Cph1d2
23.03.2004
Tycko, Robert (Laboratory of Chemical Physics, NIDDK,
Bethesda, USA)
Structure of Unfolded Proteins and Amyloid Fibrils: Experimental
Constraints from Solid State NMR 13.10.2004
Hosts: Reif B, Oschkinat H
Wüstner, Daniel (Max-Delbrück-Center, Berlin-Buch, Germany)
Transport und function of cholesterol in epi
01.12.2004
Hosts: Rosenthal W, Bienert M
Appendix
155

TECHNOLOGY TRANSFER 2003/2004
Also in 2003 and 2004, the FMP submitted patent applications
for economically promising research results, assessed
the prospects for commercial success and, where
appropriate, initiated commercialization activities. Due to
the proximity to biotech companies on the Berlin-Buch
campus and in the Berlin-Brandenburg region, excellent
conditions for the utilization of research results are provided.
To further optimize commercialization activities, the FMP
entered a contract agreement with Ascension GmbH in
Munich. Since that time Ascension has been assessing
inventions of FMP employees with regard to patents and
especially to market-relevant aspects.
The FMP currently holds eight registered or granted
patents on a total of four inventions. Applications for additional
patents have been submitted.
TECHNOLOGIETRANSFER 2003/2004
Auch in den Jahren 2003 und 2004 hat das FMP für wirtschaftlich
vielversprechende Forschungsergebnisse
Schutzrechte angemeldet, Verwertungschancen geprüft
und gegebenenfalls Maßnahmen zur kommerziellen Verwertung
ergriffen. Durch die räumliche Nähe zu biotechnologischen
Firmen auf dem Campus Berlin-Buch und in
der Region Berlin/Brandenburg sind optimale Voraussetzungen
für die Anwendung von Forschungsergebnissen
gegeben.
Um die Verwertungsaktivitäten weiter zu optimieren,
wurde im August 2004 eine Zusammenarbeit mit der
Ascenion GmbH (München) vertraglich vereinbart. Seitdem
bewertet die Ascenion Erfindungen von FMP-Mitarbeitern
hinsichtlich patent- und vor allem marktrelevanter
Gesichtspunkte.
Das FMP hält derzeit acht eingetragene oder erteilte
Schutzrechte auf insgesamt vier Erfindungen. Für weitere
Erfindungen sind Anmeldeverfahren zur Erteilung von
Schutzrechten eingeleitet.
Inventions and patents
Erfindungen und Schutzrechte
Hagen V, Bauer PJ
Als Verknüpfungsreagenzien einsetzbare dimaleinimidosubstituierte
Dihydroxyalkane und Verfahren zu deren Herstellung
DE 195 33 867 C1 (filed 24.04.1997)
EP 96938012.0 (filed 02.05.2003)
PCT/DE96/01742
US 09/043,263 (filed 15.06.1999)
priority establishing patent application: 13.09.1995
Hagen V, Kaupp UB
Neue photolabile 8-substituierte cyclische Nucleotidester,
Verfahren zu ihrer Herstellung und Verwendung
DE 195 29 025.9
EP 96927617.9-1270
WO 97/05155
priority establishing patent application: 29.07.1995
FMP-inventors in bold.

Hagen V, Kaupp B, Bendig, Wiesner B
Neue, Photolabile Cumarinylmethylester von cyclischen
Nucleotiden, Verfahren zu deren Herstellung und ihre Verwendung
DE 100 21 256 A1
PCT/EP01/03512
EP 01 929 464.4-2110
JP 2001-578448
US 01/03512
priority establishing patent application: 20.04.2000
Labudde D, Leitner D, Schubert M, Winter R, Oschkinat H,
Schmieder P
Vorrichtung und Verfahren zur Zuordnung der NMR-Signale
von Polypeptiden
DE 10144661.6-33 (filed 14.08.2003)
EP 02777009.8
PCT/EP02/09959
US 10/799,447
priority establishing patent application: 11.09.2001
Labudde D
Verfahren zur Ermittlung von Verschiebungen der Hirnareale
durch Tumorbildung und deren Darstellung auf
einem Bildschirm
DE 100 13 360.6 (filed 22.07.2004)
PCT/DE01/00955
JP 002003527899 T2
US 10/221,064
EP 01916923.4
priority establishing patent application: 09.03.2000
Maric K
Tablett für Feuchtkammer (Gebrauchsmuster)
DE 2002109.5 (filed 05.04.2001)
priority establishing application: 8.12.2000
Meyer T, Vinkemeier U
Verwendung von dimer-spezifischen Nuclear-Lokalisations-Signalpeptiden
(dsNLS) abgeleitet aus der STAT-DNA
Bindungsdomäne
DE 102 01 791.3
PCT/EP2003/000240
EP 03702426.2-2107
priority establishing patent application: 17.01.2002
Rosenthal W, Klußmann E, Oksche A
Neue Spleißvariante eines Proteinkinase A-Ankerproteins
und Verwendung dieser
DE 103 06 085.5
PCT/EP2003/09892
CA PCT/EP 2003/09892
EP 03 750 490.9-2405
JP 502327850
US 10/526.768
priority establishing patent application: 7.2.2003
Rosenthal W, Klußmann E, Hundsrucker C
Peptide zur Inhibition der Interaktion von Proteinkinase A
und Proteinkinase A-Ankerprotein
DE 10 2004 031 579.5
priority establishing patent application: 29.06.2004
Soderhäll A, Kühne R, et al.
Neue, peptidomimetische, oral verfügbare LHRH Antagonisten
mit Tetrahydrocarbazol-Grundkörper
rights sold in 2004
Siems W, Walther T, Melzig MF
Verwendung von NEP-assoziierten Molekülen zur Behandlung
von nichtimmunogenen-nichthypertensiven Zivilisationskrankheiten
DE 103 11 984.1-41
PCT/DE2004/000491
priority establishing patent application: 12.3.2003
Vinkemeier U
Neue Nucleus-Export-Signalpeptide (NES), sie enthaltende
Fusionsproteine sowie deren Verwendung
DE 100 35 867 A1
PCT/EP01/08065
EP 01954035.0
US 10/333,082
priority establishing patent application: 14.07.2000
Vinkemeier U, Meyer T
A single residue modulates Tyrosine Dephosphorylation,
oligomerization, and nuclear accumulation of Stat Transcription
factors
EP 04 09 0039.1
PCT/EP2004/001462
priority establishing patent application: 10.2.2004
Vinkemeier U, Meyer T
Verfahren zur Detektion von nukleozytoplasmatischen
Transportprozessen
DE 10 2004 046 327.1
priority establishing patent application: 20.09.2004
Vinkemeier U, Meyer T, Marg A
Hyperactive Stat molecules and their use in assays
employing gene activation
EP 04 077 629.6
priority establishing patent application: 17.09.2004
FMP-inventors in bold.
Appendix
157

STRUCTURE OF THE FORSCHUNGSINSTITUT FÜR MOLEKULARE PHARMAKOLOGIE (FMP)
Forschungsverbund
Berlin e. V.
Staff Council
Burkhard Wiesner
Structural
Biology
Protein Structure
Hartmut Oschkinat
Solution NMR
Peter Schmieder
Structural
Bioinformatics
Gerd Krause
Molecular
Modelling
Ronald Kühne
Solid State NMR
Bernd Reif
Protein Engineering
Christian Freund
Cellular Signalling
Walter Rosenthal
Protein Trafficking
Ralf Schülein
Anchored Signalling
Enno Klußmann
Cellular Imaging
Burkhard Wiesner
Molecular Cell
Physiology
Ingolf E. Blasig
Biochemical
Neurobiology
Wolf-Eberhard Siems
Cellular Signalling /
Molecular Genetics
Director
Walter Rosenthal
Molecular Genetics
Ivan Horak
Cytokine Signaling
Klaus P. Knobeloch
Molecular
Myelopoiesis
Dirk Carstanjen
Cellular Signal
Processing
Uwe Vinkemeier
Chemical
Biology
Peptide Chemistry
& Biochemistry
Michael Bienert
Peptide Synthesis
Michael Beyermann
Peptide Lipid Interaction/
Peptide Transport
Margitta Dathe
Johannes Oehlke
Peptide Biochemistry
Hartmut Berger
Mass Spectrometry
Eberhard Krause
Synthetic Organic
Biochemistry
Volker Hagen
Medicinal Chemistry
Jörg Rademann
Screening Unit
Jens Peter von Kries
Public Relations
Björn Maul
Safety Officer
Hans-Ulrich Heyne
Administration,
Technical and
Scientific Services
Administration
Thomas Ellermann
Computer Services
Thomas Jahn
Technical Services
Hans-Jürgen Mevert
Library
Michael Beyermann
Renate Peters
Animal Housing
Regina Richter
Microdialysis
Service
Regina Richter
DNA Sequencing
Service
Erhard Klauschenz

ORGANIGRAMM
Forschungsverbund
Berlin e. V.
Betriebsrat
Burkhard Wiesner
Strukturbiologie Zell-Signalling/Molekulare Genetik Chemische Biologie Verwaltung, technische
und wissenschaftliche
Dienste
Proteinstruktur
Hartmut Oschkinat
Lösungs-NMR
Peter Schmieder
Strukturelle
Bioinformatik
Gerd Krause
Molekulares Modelling
Ronald Kühne
Festkörper-NMR
Bernd Reif
Protein Engineering
Christian Freund
Zell-Signalling
Walter Rosenthal
Protein-Trafficing
Ralf Schülein
Anchored Signalling
Enno Klußmann
Zell-Imaging
Burkhard Wiesner
Molekulare
Zellphysiologie
Ingolf E. Blasig
Biochemische
Neurobiologie
Wolf-Eberhard Siems
Direktor
Walter Rosenthal
Molekulare Genetik
Ivan Horak
Zytokin-Signalling
Klaus P. Knobeloch
Molekulare
Myelopoese
Dirk Carstanjen
Zelluläre
Signalverarbeitung
Uwe Vinkemeier
Peptidchemie &
Biochemie
Michael Bienert
Peptidsynthese
Michael Beyermann
Peptid-Lipid-Interaktion/Peptidtransport
Margitta Dathe
Johannes Oehlke
Peptidbiochemie
Hartmut Berger
Massenspektrometrie
Eberhard Krause
Synthetische Organische
Biochemie
Volker Hagen
Medizinische Chemie
Jörg Rademann
Screening-Unit
Jens Peter von Kries
Öffentlichkeitsarbeit
Björn Maul
Arbeitssicherheit
Hans-Ulrich Heyne
Verwaltung
Thomas Ellermann
Computer-Service
Thomas Jahn
Technischer Service
Hans-Jürgen Mevert
Bibliothek
Michael Beyermann
Renate Peters
Tierhaltung
Regina Richter
Mikrodialyseservice
Regina Richter
DNA-Sequenzierservice
Erhard Klauschenz
Structure of the FMP
159

INDEX
A
Al-Gharabli, Samer ............................................................. 90
Alken, Martina ..................................................................... 40
Andreeva, Anna Y. ............................................................... 49
Antonenko, Yuri ................................................................... 57
Appelt, Christian ............................................................ 16, 18
Ayuyan, Artem ..................................................................... 57
B
Ball, Linda ............................................................................. 13
Balling, Rudolf ........................................................................ 2
Bárány-Wallje, Elsa ............................................................ 76
Barth, Michael ..................................................................... 90
Bauer, Jörg ........................................................................... 90
Becker, Matthias ................................................................. 55
Beck-Sickinger, Annette G. ................................................. 2
Begitt, Andreas .................................................................... 66
Benedict, Melanie ............................................................... 63
Ben-Slimane, Uta ................................................................ 33
Berger, Hartmut ............................................................. 70, 80
Beyermann, Michael .................................................... 70, 72
Bienert, Michael ................................................... 70, 75, 76
Blasig, Ingolf ........................................................................ 49
Blasig, Rosel ........................................................................ 64
Blick, Helmut ...................................................................... 101
Blum, Christopher ............................................................... 43
Bohne, Kerstin ..................................................................... 64
Boisguerin, Prisca ............................................................... 13
Boldt, Liane .......................................................................... 60
Brauße, Kerstin ................................................................. 107
Bräutigam, Matthias ............................................................. 2
Brockmann, Christoph ........................................... 13, 25, 26
Buschner, Michael ............................................................ 101
C
Carstanjen, Dirk ................................................................... 62
Cartier, Regis ........................................................................ 66
Castellani, Federica ............................................................ 13
Chen, Zhongjing ................................................................... 29
Chernogolov, Alex ............................................................... 29
Chevelkov, Veniamin ........................................................... 29
Coin, Irene ............................................................................ 74
D
Dasari, Muralidhar .............................................................. 29
Dathe, Margitta ....................................................... 70, 75, 76
Dekowski, Brigitte ............................................................... 87
Diehl, Anne ........................................................................... 13
Djurica, Maja ....................................................................... 63
Donalies, Ute ........................................................................ 40
Dreissigacker, Marianne ................................................... 70
E
Ehrlich, Angelika ..................................................... 74, 75, 95
Eichhorst, Jenny .................................................................. 48
Eilemann, Barbara .............................................................. 50
Eisenmenger, Frank ...................................................... 24, 26
El-Dashan, Adeeb ............................................................... 90
Ellermann, Thomas ........................................................... 106
Erdmann, Christoph ............................................................ 95
Evers, Heide ......................................................................... 14
F
Fälber, Katja ......................................................................... 13
Fechner, Klaus ..................................................................... 82
Fink, Uwe ...............................................................................29
Flinders, Jeremy .................................................................. 13
Fossi, Michele ...................................................................... 13
Freund, Christian ........................................................... 31, 32
G
Geelhaar, Andrea ................................................................ 43
Geißler, Daniel ..................................................................... 87
Georgi, Monika .................................................................... 82
Gomoll, Michael .................................................................. 43
Göritz, Petra ....................................................................... 101
Griesinger, Christian ............................................................. 2
H
Hackel, Uwe ....................................................................... 101
Hagen, Volker ................................................................. 70, 86
Hahn, Janina .................................................................. 16, 18
Handel, Lilo ........................................................................... 14
Hartmann, Gislinde ............................................................. 50
Haseloff, Reiner F. ............................................................... 49
Hausbeck, Dana ................................................................ 107
Heine, Markus ..................................................................... 50
Heinrich, Nadja ................................................................... 82
Heinze, Matthias ........................................................... 31, 33
Henn, Volker ......................................................................... 43
Hermann, Ingrid ................................................................. 108
Heuer, Katja .................................................................... 32, 33
Heyne, Alexander .............................................................. 108
Heyne, Hans-Ulrich ........................................................... 101
Hiller, Matthias ..................................................................... 13

Hologne, Maggy ...................................................................29
Holtmann, Henrik ................................................................. 13
Horak, Ivan ........................................................................... 36
Hübel, Stefan ....................................................................... 26
Hundsrucker, Christian ....................................................... 43
J
Jahn, Reinhard ...................................................................... 2
Jahn, Thomas .................................................................... 108
Jehle, Stefan ........................................................................ 14
Joost, Hans-Georg ................................................................ 2
Joshi, Mangesh ................................................................... 13
K
Kallies, Axel .......................................................................... 63
Kahlich, Bettina ................................................................... 55
Keller, Sandro ................................................................ 76, 78
Kiesling, Alexandra ............................................................. 36
Kirsch, Jenny ....................................................................... 50
Kisser, Agnes ....................................................................... 60
Klauschenz, Erhard ........................................................... 101
Kleinau, Gunnar ............................................................. 20, 23
Klemm, Clementine .............................................................. 85
Klemm, Janet ....................................................................... 64
Klose, Annerose .................................................................. 74
Klose, Jana............................................................................ 74
Klussmann, Enno ................................................................. 42
Knobeloch, Klaus-Peter ..................................................... 59
Köhler, Christian .................................................................. 13
Königsmann, Jessica ...........................................................63
Kofler, Michael ..............................................................31, 33
Kotzur, Nico............................................................................87
Krabben, Ludwig .................................................................. 13
Krätke, Oliver .........................................................................74
Krause, Dagmar.....................................................................74
Krause, Eberhard ........................................................... 70, 83
Krause, Gerd......................................................................... 20
Krause, Winfried .................................................................. 55
Kries, Jens-Peter von................................................ 3, 70, 92
Krylova, Oxana O.................................................................. 57
Kühne, Ronald................................................................. 24, 25
Kummerow, Mandy ............................................................. 66
L
Labudde, Dirk ....................................................................... 13
Lättig, Jens...................................................................... 20, 23
Lamer, Stephanie ................................................................. 85
Lange, Vivien ........................................................................ 14
Lassowski, Birgit ................................................................. 50
Lauterjung, Ulrike .............................................................. 104
Lechler, Ralf ......................................................................... 87
Leidert, Martina ................................................................... 14
Lentz, Alexander................................................................... 57
Leitner, Dietmar ................................................................... 13
Lerch, Heidi .......................................................................... 85
Liesenfeld, Oliver ................................................................. 76
Lojek, Eva ............................................................................ 101
Lödige, Inga .......................................................................... 66
Lorberg, Dörte ...................................................................... 50
Lorenz, Dorothea ........................................................... 43, 48
M
Mac, Thien-Thi .................................................................... 14
Manks, Silvia ...................................................................... 107
Marg, Andreas ..................................................................... 66
Margania, Valentina ........................................................... 57
Maul, Björn ................................................................. 102, 103
McSorley, Theresa .............................................................. 43
Meier, Franziska .................................................................. 90
Meissner, Torsten ................................................................ 66
Melchior, Frauke ................................................................... 2
Messing, Claudia ............................................................... 107
Mevert, Jürgen....................................................................101
Meyer, Stephanie ................................................................ 66
Meyer, Thomas .................................................................... 66
Michl, Dagmar ..................................................................... 40
Mikoteit, Kerstin .................................................................. 50
Mohs, Barbara ................................................................... 101
Mollajew, Rustam ................................................................ 57
Motzny, Katrin ................................................................ 31, 33
Müller, Sebastian L. ................................................ 20, 23, 50
Muschter, Antje ................................................................. 100
N
Narayanan, Saravanakumar ............................................. 29
Nedvetsky, Pavel ................................................................. 43
Niehage, Christian .............................................................. 50
Niendorf, Sandra ................................................................. 60
Nikolenko, Heike ................................................................. 78
Index
161

O
Oczko, Brunhilde ................................................................. 48
Oehlke, Johannes ......................................................... 70, 75
Oschkinat, Hartmut ....................................................... 10, 12
Osiak, Anna .......................................................................... 60
Otto, Christel ...................................................................... 107
Oueslati, Morad ................................................................... 40
P
Pahlke, Doreen .................................................................... 14
Pankow, Kristin .................................................................... 55
Pankow, Rüdiger ................................................................. 64
Panzer, Holger .................................................................... 101
Passow, Josephine ........................................................... 107
Perepellichenko, Ludmila ................................................... 90
Peters, Renate ................................................................... 101
Petschick, Heidemarie ....................................................... 36
Piontek, Jörg ........................................................................ 49
Piotukh, Kirill .................................................................. 31, 33
Pires, Ricardo ...................................................................... 13
Pisarz, Barbara .................................................................... 74
Pisarz, Hans-Werner ........................................................ 108
Pohl, Peter ............................................................................ 57
Poliakov, Ilja ......................................................................... 14
Prigge, Matthias .................................................................. 57
Pritz, Stephan ....................................................................... 74
R
Rademann, Jörg ........................................................ 3, 70, 89
Reif, Bernd ............................................................................ 28
Rehbein, Kristina ................................................................. 14
Richter, Regina M. ....................................................... 98, 101
Ringling, Martina ................................................................. 48
Rosenthal, Walter ............................................................ 6, 36
Rossum, Barth van .............................................................. 13
Roswadowski, Inga ............................................................. 50
Rötzschke, Olaf .................................................................... 25
Rückert, Christine ............................................................... 49
Rutz, Claudia ........................................................................ 40
S
Santamaria, Katja ................................................................ 43
Saparov, Sapar M. .............................................................. 57
Sauer, Ines ..................................................................... 76, 78
Scharnagel, Hubert ............................................................. 76
Schlegel, Brigitte ........................................................... 16, 18
Schmidt, Antje ..................................................................... 46
Schmieder, Peter ................................................................. 16
Schmikale, Bernhard .......................................................... 74
Schneeweiß, Ulrike ............................................................ 33
Schülein, Ralf ....................................................................... 33
Schümann, Michael ............................................................ 85
Schumacher, Gabriele ...................................................... 107
Schumann, Björn ............................................................... 108
Schrey, Anna .................................................................. 24, 26
Schröder, Nikolaj ................................................................. 14
Schulz, Katrin ....................................................................... 50
Schuster, Ariane .......................................................... 50, 152
Selfe, Joanna ....................................................................... 63
Serowy, Steffen ............................................................. 57, 76
Shan, Ying ............................................................................ 66
Siems, Wolf-Eberhard ........................................................ 52
Singh Bal, Manjot ................................................................ 50
Söderhäll, Arvid ....................................................... 24, 25, 26
Sokolenko, Elena ................................................................. 57
Sokolov, Valerij .................................................................... 57
Soukenik, Michael .............................................................. 14
Souza, Christina de ............................................................. 57
Sperling, Birgit ................................................................... 107
Stefan, Eduard ..................................................................... 43
Steuer, Andrea ..................................................................... 10
Strauss, Holger .............................................................. 16, 18
Sun, Xiaoou .......................................................................... 55
Sylvester, Marc ............................................................. 32, 33
T
Tasadaque Ali Shah, Syed ................................................. 90
Tenz, Kareen ......................................................................... 85
Thielen, Anja ........................................................................ 40
Thiemke, Katharina ................................................. 31, 32, 33
Thakur, Mina ........................................................................ 64
Tremmel, Sandra ................................................................. 74
Tsunoda, Satoshi ................................................................. 56
U
Uryga-Polowy, Viviane ....................................................... 90
Uschner, Michael .............................................................. 101
Utepbergenov, Darkhan ..................................................... 49
V
Vargas, Carolyn ................................................................... 14
Venta, Nicola ........................................................................ 66
Vinkemeier, Uwe .................................................................. 65
Vogelreiter, Gabriela ..................................................... 78, 82
W
Waldmann, Herbert ............................................................... 2
Walter, Juliane ..................................................................... 50
Weber, Viola ......................................................................... 43
Weik, Steffen ........................................................................ 90
Weisgraber, Karl .................................................................. 76
Wendt, Stefanie ................................................................. 101
Wessolowki, Axel ................................................................ 78
Wichard, Jörg ...................................................................... 26

Wiedemann, Urs ...................................................... 14, 20, 23
Wiesner, Burkhard ........................................................ 46, 75
Wietfeld, Doreen ................................................................. 74
Wietstruck, Markus ............................................................ 60
Winkler, Lars ........................................................................ 50
Wolf, Constanze ................................................................... 50
Wolf, Yvonne .................................................................. 75, 78
Wolff, Christian .................................................................. 100
Z
Zimmermann, Jürgen ................................................... 32, 33
Zuleger, Nikolaj ............................................................ 50, 145
Index
163

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165