International Protease Network

Institute for Glycomics
Griffith University
PMB 50 Gold Coast Mail Centre, Queensland 9726
Email: h.blanchard@griffith.edu.au
Tel: +61-7-555-27023
Fax: +61-7-555-28098
Homepage:

Research

We are interested in structure-based drug design approaches and as one direction with application within protease research. We are using protein X-ray crystallography as the main technique for providing atomic resolution three-dimensional structural information. This approach provides the exact details of the protein active-site region, the specific protein-ligand interactions and of the local water structure. This information is important in ascertaining characteristics of the binding pocket and gives the specific ligand interactions important in binding and recognition. Using structure analysis and molecular modelling, and in combination with ligand-binding assays, we are focused on design of molecules that maximise favourable interactions with the protein thereby ultimately leading to the design of potent therapeutics.
On the calpain system, a calcium-regulated cytoplasmic protease with roles in signal transduction, apoptosis, cell-cycle regulation and cytoskeletal reorganisation, we contributed to the understanding of calcium-regulation of protease activity. We determined the structure of the calcium-binding domain in the presence and absence of calcium providing the first three-dimensional structural information on the calpain system. This gave insight into calpain drug-design preventing protease activation via disruption of the action of calcium-induced structural changes as an alternative to targeting the cysteine protease active site itself. Another area of interest are cysteine proteases involved in apoptosis (programmed cell-death), a normal physiological process which when uncontrolled can lead to neurological diseases and cancer. The apoptotic process is a cascade of events and we focused on drug-design of caspase-8, determining via X-ray crystallography, the first structure of this initiator enzyme of apoptosis.

Collaborations

Publications

N.C.J. Strynadka, S.E. Jensen, K. Johns, H. Blanchard, M. Page, J-M. Frere, and M.N.G James. Structural and kinetic characterisation of a novel proteinaceous beta-lactamase inhibitor from Streptomyces Clavuligerus. (1994) Nature 368 No. 6472 657-660.

H. Blanchard and M.N.G James. Crystallographic re-investigation into the structure of Streptomyces griseus proteinase A reveals an acyl-enzyme. (1994) Journal of Molecular Biology 241 No. 4 574-587.

H. Blanchard, P. Grochulski, Y. Li, J. S. C. Arthur, P.L. Davies, J.S. Elce and M. Cygler. Structure of a calpain Ca2+-binding domain reveals a novel EF-hand and Ca2+-induced conformational changes. (1997) Nature Struct. Biol. 4 532-538.

H. Blanchard., L. Kodandapani., P.R.E, Mittl., S. Di Marco., J.F. Krebs., J.C. Wu., K.J. Tomaselli., M.G. Grutter. The three-dimensional structure of caspase-8, an initiator enzyme in apoptosis. (1999) Structure Fold. Des. 7 : 1125-1133.

H. Blanchard, M. Donepudi, M. Tschopp, L. Kodandapani, J.C. Wu, M.G. Grutter. J Mol Biol. Sep 8; 302(1) :9-16 (2000) Caspase-8 specificity probed at the subsite S4: crystal structure of the caspase-ZDEVD-cho complex.