Biophysical characterisation of proteins and enzymes
Biophysical characterisation
We employ a wide variety of biophysical characterisation techniques in addition to traditional enzyme kinetic assays including isothermal titration calorimetry (ITC), fluorescence methods such as fluorescence anisotropy and analytical ultracentrifugation. These techniques have been applied to a wide variety of binding systems including canonical phosphotyrosine-binding domains such as SH2 domains as well as protein targets related to cancer progression.
For characterisation of enzyme activity, we use a variety of approaches in addition to enzyme activity assays including characterisation of ligand binding using ITC, structural characterisation using X-ray crystallography, MS and NMR and site-directed mutagenesis. We are interested in the function of all enzymes but have particular experience of working with the following:
- Pyruvate, orthophosphate dikinase regulatory protein
- AICAR transformylase
- Aspartate α-decarboxylase
- ADC activating protein PanZ
- Homoprotocatechuate dioxygenase
- Sortase
- Sirohaem chelatase
Recent publications in this area
The Structure of the PanD/PanZ Protein Complex Reveals Negative Feedback Regulation of Pantothenate Biosynthesis by Coenzyme A
Diana C.F. Monteiro, Vijay Patel, Christopher P. Bartlett, Shingo Nozaki, Thomas D. Grant, James A. Gowdy, Gary S. Thompson, Arnout P. Kalverda, Edward H. Snell, Hironori Niki, Arwen R. Pearson & Michael E. Webb Chem. Biol. (2015) 22 492-503
Formation of a heterooctameric complex between ADC and its cognate activating factor, PanZ, is CoA-dependent Diana C.F. Monteiro, Michael D Rugen, Dale Shepherd, Shingo Nozaki, Niki Hironori and Michael E Webb Biochem. Biophys. Res. Commun. (2012) 426 350-355
Characterisation of the evolutionarily conserved iron sulphur cluster of sirohydrochlorin ferrochelatase from Arabidopsis thaliana Kaushik Saha, Michael E Webb, Stephen EJ Rigby, Helen K Leech, Martin J Warren and Alison G Smith Biochem. J. (2012) 444 227-237