NMR Spectroscopy
Prof J P Waltho - Gibson Chair in Biophysics |
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| Our laboratory focuses on the use of high field NMR spectroscopy to determine the structure and function of proteins and how they fold from their fully unfolded states. Our studies address both fundamental aspects of protein biophysics and dynamics, and the investigation of the biomedical targets and their inhibition. |
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Recent highlights include the structure determination and characterisation of the solution dynamics of the human intracellular cysteine proteinase inhibitor stefin A. Proteins of this family inhibit enzymes central to the invasion of the body by foreign organisms (e.g. trypanosomes that cause African Sleeping sickness) and the entry of metastatic cancer cells into new tissues. In addition, the absence of the protein stefin B was recently the first identified genetic cause of epilepsy. Structure and dynamic measurements of stefins provide insights into means of developing small molecule pharmaceuticals that mimic the protective function of this class of proteins. |
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Understanding how proteins fold is both a major goal from a viewpoint of fundamental biochemistry, and is of growing biomedical importance owing its implication in a variety of neurodegenerative diseases. Diseases ranging from Alzheimer’s, through amyloid angiopathy to the prion diseases, CJD and BSE, appear to utilise partially and misfolded states of proteins. We have shown how NMR can be used to determine structural and dynamic information of such states, which provide a basis for identifying where and how to interrupt amyloidogenesis before the onset of neurodegeneration. We focus on three proteins in this regard, cystatin C, human prion protein and phosphoglycerate kinase (PGK). |
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Figure Legend - A ribbon representation of the structure of the N-terminal domain of PGK when it is fully folded with the regions in which hydrogen bonds are formed in its major kinetic folding intermediate marked with spheres. The radius of the spheres represents the strengths of the hydrogen bonds of the partially folded protein. |
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Selected PublicationsStructure of a kinetic protein folding intermediate by equilibrium amide exchange. LLP Hosszu, CJ Craven, MJ Parker, M Lorch, J Spencer, AR Clarke and JP Waltho, Nature Structural Biology (1997) 4 801-804. |
| Topology, sequence evolution and folding dynamics of an immunoglobulin domain. MJ Parker, CE Dempsey, LLP Hosszu, JP Waltho and AR Clarke, Nature Structural Biology (1998) 5 194-198. |
| Reversible inter-conversion of monomeric human prion protein between native and fibrilogenic conformations. GS Jackson, LLP Hosszu, A Power, AF Hill, J Kenney, H Saibil, CJ Craven, JP Waltho, AR Clarke and J Collinge, Science (1999) 283 1935-1937. |
| The major transition state in protein folding need not involve the immobilisation of side chains. RA Staniforth, JLE Dean, Q Zhong, AR Clarke, E Zerovnik and JP Waltho, Proc Nat. Acad. Sci. USA (2000) 97 5790-5795. |
| Location and properties of metal-binding sites on the human prion protein. Jackson GS, Murray I, Hosszu LLP, Gibbs N, Waltho JP, Clarke AR, Collinge J, Proc Nat. Acad. Sci. USA (2001) 98 8531-8535 |
| Three-dimensional domain swapping in the folded and molten-globule states of cystatins, an amyloid-forming structural superfamily. Staniforth RA, Giannini S, Higgins LD, Conroy MJ, Hounslow AM, Jerala R, Craven CJ, Waltho JP, EMBO J. (2001) 20 4774-4781 |
| Residue 129 polymorphism in human prion protein does not confersusceptibility to Creutzfeldt-Jakob disease by altering the structure orglobal stability of PrPC. Hosszu LLP, Jackson GS, Trevitt CR, Jones S, Batchelor M, Bhelt D, Prodromidou K, Clarke AR, Waltho JP, Collinge J J. Biol. Chem. (2004) 279 28515-28521 |
| Enhanced ligand affinity for receptors in which components of the binding site are independently mobile. Trevitt CR, Craven CJ, Milanesi L, Syson K, Mattinen ML, Perkins J, Annila A, Hunter CA, Waltho JP Chem. Biol. (2005) 12 89-97. |
| Determinants of the endosomal localization of sorting nexin 1. Zhong Q, Watson MJ, Lazar CS, Hounslow AM, Waltho JP, Gill GN Mol. Biol. Cell (2005) 16 2049-2057 |
| Solution structure of the helicase interaction domain of the primase DnaG: a model for helicase activation. Syson K, Thirlway J, Hounslow AM, Soultanas P, Waltho JP Structure (2005) 13 609-616 |