2010 Annual Report - Institute for Molecular Bioscience - University ...

imb annual report 2010
60
JENNY MARTIN
PROTEIN STRUCTURE AND DRUG DESIGN
OUR GOAL IS TO BETTER UNDERSTAND
the role of proteins in disease and to
develop novel drugs targeting diseasecausing
proteins. We use a range of
biochemical and biophysical techniques
to investigate the structure, function and
interactions of proteins, with a particular
emphasis on high-throughput protein
crystallography and structure-based
approaches for inhibitor design.
A major outcome over the past few
years has been the tremendous advance
in our understanding of the regulation
of SNARE proteins involved in insulinstimulated
trafficking of the GLUT4 glucose
transporter. This process, which is critical
to the regulation of blood glucose levels, is
affected in Type II Diabetes. We had shown
in collaboration with Professor David
James (Garvan Institute), that the regulatory
protein Munc18c may stimulate delivery
of GLUT4 vesicles to the cell membrane,
through binding to a short N-terminal
peptide of the SNARE syntaxin4 protein
(Latham et al. Traffic 2006; Hu et al. Proc
Natl Acad Sci USA 2007). Very recently we
identified that the Syntaxin N-peptide does
not discriminate well between cognate
and non-cognate Munc18 proteins and
that its role may instead be to induce a
conformational change in Munc18 proteins
(Hu and Christie et al. Proc Natl Acad Sci
USA 2010).
Our long-running interest in bacterial redox
folding factors led to the determination
of the crystal structure of a key virulence
factor, DsbA from Pseudomonas
aeruginosa (Shouldice et al., AntiOx Redox
Signal 2009) using data measured at
the Australian Synchrotron. We are now
focusing our attention on developing
inhibitors of several DsbA proteins as
potential antibacterial agents (Heras et al.
Nature Rev Microbiol 2009), using a range
of techniques. Collaborations have been
established with Associate Professor Martin
Scanlon at Monash and Professors David
Fairlie, Matt Cooper and Mark Schembri at
UQ. This research led to the award of an
ARC Australian Laureate Fellowship that
was taken up in late 2009.
We have successfully applied a fragmentscreening
approach to PNMT, the
adrenaline synthesising enzyme, using the
automated UQ ROCX diffraction facility
(Drinkwater et al., Biochem J 2010). More
than 140 PNMT crystals were used to
screen ~380 drug-like fragments: 12 hits
were identified and confirmed by isothermal
calorimetry. Six elaborated compounds
were designed and synthesised (in
collaboration with Gary Grunewald, Kansas
U) and shown to inhibit enzyme activity (in
collaboration with Michael McLeish, Indiana
U).
RESEARCH PROJECTS
• Structure, function and interactions of
proteins associated with insulin action
• Structure, function and inhibition of
redox folding factors involved in disease
• Novel inflammation drug targets using
high-throughput structure approaches
• Structure, function and inhibition of
transferase enzymes involved in disease
KEY PUBLICATIONS
Drinkwater, N. et al. (Martin, J.L. listed
as senior author) (2010). Fragmentscreening
by X-ray crystallography, mass
spectrometry and isothermal titration
calorimetry to identify PNMT inhibitors.
Biochemical Journal 431: 51-61.
Hu, S.-H. et al. (Martin, J.L. listed as
senior author) (2010). Possible roles for
Munc18-1 domain 3a and Syntaxin1
N-peptide and C-terminal anchor in
SNARE complex formation. ] Proceedings
of the National Academy of Sciences USA
Epub December 30.
Shouldice, S.R. et al. (Martin, J.L. listed
as senior author) (2010). Characterisation
of the DsbA oxidative folding catalyst
from Pseudomonas aeruginosa reveals a
highly oxidizing protein that binds small
molecules. Antioxidants & Redox Signaling
12: 921-931. (Front cover)
Heras, B. et al. (Martin, J.L. listed as
senior author) (2009). Dsb proteins and
bacterial pathogenicity. Nature Reviews
Microbiology 7: 215-225.
Hu, S.-H., Latham, C.F., Gee, C.L.,
James, D.E., and Martin, J.L. (2007).
Structure of the Munc18c/Syntaxin4
N-peptide complex defines universal
features of the N-peptide binding mode of
SM proteins. Proceedings of the National
Academy of Sciences USA 104: 8773-
8778.
Gruber, C., Cemazar, M., Heras, B.,
Martin, J.L., and Craik, D.J. (2006).
Protein disulfide isomerase: The structure
of oxidative folding. Trends in Biochemical
Sciences 31: 455-464.
LAB MEMBERS
Senior Research Officers: Dr Begoña
Heras, Dr Shu-Hong Hu
Research Officers: Dr Andrew Whitten
(NHMRC Fellow), Dr Stephen Shouldice
(UQ Postdoctoral Fellow), Dr Gordon King,
Dr Maria Halili, Dr Karl-Fredrik Lindahl, Dr
Morten Groftehauge, Dr Mathieu Coincon
UQ ROCX Diffraction Facility Manager:
Karl Byriel
Research Assistants: Russell Jarrott,
Stephanie Tay, Fabian Kurth
PhD Students: Michelle Christie, Kevin
Chen, Asma Rehman, Patricia Walden,
Wilko Duprez
Undergraduate Student: Heather Nutt
Visiting Student: Pooja Sharma (Monash
University)