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Dr Stephen P Bottomley
Dept. of Biochemistry and Molecular Biology
Monash University
Clayton, Vic 3800
Email: steve.Bottomley@med.monash.edu.au
Tel: +61-3-99053703
Fax: +61-7-99054699
Homepage: http://www.med.monash.edu.au/biochem/staff/bottomley.html
| Research |
We are interested in the role of the human protease inhibitors from the serpin superfamily. We have used biochemical, biophysical and structural approaches to study their form and function. In particular we have used rapid kinetics and fluorescent labeling techniques to follow the conformational changes involved in forming the serpin-protease complex. This work involved the development of novel kinetic mechanisms and mathematical models. We have also used this information along with detailed bioinformatic analysis to develop novel serpin molecules with defined specificity and high stability, which may prove useful in the treatment of a number of disorders where unwanted proteolysis occurs.
Recently, we have also studied how these protease inhibitors fold and misfold. When the serpin misfolds it is retained at its site of synthesis instead of being secreted. This leads to a lack of circulating proteinase inhibitor which leads to excessive proteolysis and tissue damage. In addition the aggregation of these proteins during their folding leads to cellular damage and a range of devastating human diseases including emphysema, liver cirrhosis, thrombosis, and dementia. Our work was the first to kinetically and structurally characterise these misfolding events and identify key steps in the process. A particular highlight was the determination of the structure of a serpin polymer. Recently, we have also developed high-throughput assays which are allowing us to screen large chemical libraries for the development of potential therapeutics.
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| Collaborations |
Dr Bernard Le Bonniec, INSERM, U428, Universite Paris V, Faculte de Pharmacie 4 Avenue de lObservatoire, 75270 Paris Cedex 06, France
Dr Dimitri Gilis, Ingenierie Biomoleculaire, Universite Libre de Bruxelles, CP 165/64, 50 avenue Roosevelt, 1050 Brussels, Belgium
Dr Michael Gore, Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, Hants. SO16 7PX, UK
Prof. David Lomas, Department of Haematology, University of Cambridge, Wellcome Trust Centre for Molecular Mechanisms in Disease, Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, Hills Road, Cambridge, CB2 2XY, UK
Prof. Henry Paulson, Department of Neurology, University of Iowa College of Medicine, Iowa City, IA 52252, USA
Prof. Harvey Rubin, Department of Medicine, University of Pennsylvania, Philadelphia 19104, USA
A/Prof John Carver, Dept. of Chemistry, University of Wollongong
Dr Robert Pike, Dr James Whisstock and Dr Phillip Bird, Dept. of Biochemistry and Molecular Biology, Monash University
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| Publications |
Devlin, G.L., Chow, M.K., Howlett, G and Bottomley, S.P. Acid Denaturation of a1-Antitrypsin:Characterization of a Novel Mechanism of Serpin Polymerization. (2002) J. Mol. Biol 324, 859-870
Devlin, G, L., Parfrey, H., Tew, D.J., Lomas, D.A. and Bottomley, S.P. (2001) Prevention of polymerization of M and Z a1-antitrypsin (a1-AT) with Trimethylamine N-Oxide: Implications for the treatment of a1-AT deficiency. Am. J. Respir. Cell Mol. Biol. 24, 727-732.
Ludeman, J.P., Whisstock, J.C., Hopkins, P.C., Le Bonniec, B.F. and Bottomley, S.P. (2001) The structure of a serpin-enzyme complex probed by cysteine substitutions and fluorescence spectroscopy. Biophys. J. 80, 491-497.
James, E.L., Whisstock, J.C., Gore M.G. and Bottomley, S.P. (1999) Probing the unfolding pathway of a1-Antitrypsin. J. Biol. Chem. 274, 9482 - 9488.
Dunstone, M.A., Dai, W., Whisstock, J.C., Rossjohn, J., Pike, R.N., Feil, S.C., Le Bonniec, B., Parker M.W. and Bottomley, S.P. (2000) Cleaved Antitrypsin Polymers at Atomic Resolution Protein Science 9, 429 - 432.
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