Thomas Jowitt, BSc MSc PhD
The biomolecular analysis core facility is much more than a room full of instruments. We are a technical and eruditional resource with the aim of aiding research programs, teaching, technical and scientific development and bringing together a wider scientific community who share similar biophysical questions.
Two of the staff members have a long history of association with the Wellcome Centre, Marj Howard and Tom Jowitt, and as such methodological development of the instruments has often been optimised for the demanding requirements of large matrix-associated molecules. In particular, the development of hydrodynamic modelling techniques has helped solve the solution structure of several matrix macromolecules which have previously been difficult to predict
The facility can be broadly categorised into the two complimentary themes of structural hydrodynamics and interactions. These two themes are highly complex and often difficult to appreciate, but we have developed the facility to make the investigation of molecular structure, conformation, dynamics and interactions as transparent as possible.
We have optimised the development of hydrodynamic bead models and constrained rigid-body dynamics of oligomeric assemblies which allows us to render low resolution structural information of multi-domain and oligomeric species. We are now in a position where we can model dynamics of either singular or multi-domain molecules and compare the simulations with real-time hydrodynamic data such as can be seen here in the tail-flexibility simulation of Cub-c which is in collaboration with the Day lab.
We are also developing a range of functionalised lipid bi layers and tethered vesicles which we are using as a support for protein immobilisation. These surfaces are generally less reactive than conventional silicon dioxide or carbohydrate functionalised supports and therefore allow us to investigate complex interaction kinetics of difficult to work with proteins. We are also able to optimise the surface density of protein immobilisation and therefore accurately regulate the interactions at the surface. Using Farfield interferometry and quartz crystal microbalance techniques we can probe the surfaces to reveal accurate thickness, density and mass measurements. We can now orient our molecules in such a way as to gain detailed structural information and insights into molecular associations with small molecules.
We can also monitor the birefringence of these anisotropic layers and so determine if the molecules are integrating within the layers, therefore disturbing the order within the lipid layers.This will occur with insertion of a membrane protein or ion channel or with membrane perturbation caused by antimicrobial peptides. Taken together these techniques allow us to build accurate and detailed information on the solution characteristics of molecules, how they interact with their partners and whether these interactions are characterised by structural and environmental consequences
Tel: +44 (0) 161 306 5176
Recent key publications
Berry, R., Jowitt, T. A., Ferrand, J., Roessle, M., Grossmann, J. G., Canty-Laird, E. G., Kammerer, R. A., Kadler, K. E., and Baldock, C. (2009) Role of dimerization and substrate exclusion in the regulation of bone morphogenetic protein-1 and mammalian tolloid. Proc Natl Acad Sci U S A. 106, 8561-6.
Jowitt, T. A., Murdoch, A. D., Baldock, C., Berry, R., Day, J. M., and Hardingham, T. E. (2010) Order within disorder: Aggrecan chondroitin sulphate-attachment region provides new structural insights into protein sequences classified as disordered. Proteins. 78, 3317-27.
Fresquet, M., Jowitt, T. A., Stephen, L. A., Ylostalo, J., and Briggs, M. D. (2010) Matrilin-1 A-domains: structural and functional investigations reveal insights into their role in cartilage ECM assembly. J Biol Chem. 285, 34048-61.