walthoj

Prof Jon Waltho
Gibson Chair in Biophysics

Room: B108
0114 222 2717
j.waltho@sheffield.ac.uk

General

Career History

  • 2009: Awarded the Gibson Chair of Biophysics
  • 2008: Professorial Appointment in Faculty of Life Sciences, University of Manchester
  • 1999: Awarded a Personal Chair in Molecular Biology and Biotechnology
  • 1997: Promoted to a Readership
  • 1996: Awarded a Research Fellowship by the Lister Institute of Preventive Medicine
  • 1996: Promoted to a Senior Lectureship
  • 1990: Awarded a Krebs Institute Lectureship in the Department of Molecular Biology and Biotechnology, University of Sheffield
  • 1987 - 90: Postdoctoral research assistant, with Dr P. E. Wright, Department of Molecular Biology, The Scripps Research Institute, La Jolla, California
  • 1986 - 87: Postdoctoral research assistant, with Dr J. G. Vinter, SmithKline French Research
  • 1983 - 86: Postgraduate studentship, SERC funded, with Dr D. H. Williams in the University Chemical Laboratory and Emmanual College, Cambridge
  • 1980 - 83: B.Sc. Honours degree in Chemistry, Class I, at Grey College, Durham

Research Keywords

Structural biology, NMR, enzymology, kinetics, protein folding, prions

Research

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.

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.

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).

fig1

Figure 1: 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.

Teaching

Level 3 Modules

PhD Opportunities

I welcome applications from self-funded prospective home and international PhD students; see examples of possible projects below.

You can apply for a PhD position in MBB here.

Contact me at j.waltho@sheffield.ac.uk for further information.



Biophysics of enzyme-catalyzed phosphoryl transfer

Using a combination of multinuclear NMR spectroscopy, high resolution X-ray crystallography, synthetic chemistry and computational chemistry we have explored the conformational behaviour of enzymes under a very wide range of conditions. Recently, our group has focused on enzymes that catalyse the transfer of phosphoryl groups, where the non-catalysed reactions can be among the slowest known for a physiological process: phosphate monoesters, for example, have calculated lifetimes to spontaneous hydrolysis of up to 1012 years. We are using a range of phosphoryl transfer enzymes to illustrate what contributes to the very high levels of catalysis achieved by these enzymes. The introduction of metal fluoride species to mimic the ground states and the transition state of the transferring phosphate group has allowed us to dissect the steps involved in catalysis, and to dissect electronic contributions made by the enzymes. In particular, these enzymes illustrate how the enzyme tightly controls charge distribution in the close vicinity of the transferring phosphate, and how the near transition state complex conformation reacts to modulation of these charges. Now, we will extend these observations to detail how phosphoryl transfer enzymes control domain closure, and how the distribution of protons plays key roles in controlling catalysis.









































Publications

Journal articles

Chapters

  • Waltho JP, Bowler MW, Cliff MJ & Vas M (2013) Architecture of the active site and the importance of charge balance in catalysis, Phosphoglycerate Kinase: A Hinge-Bending Enzyme (pp. 203-210).

Conference proceedings papers

  • Finger LD, Bennet IA, Hounslow A, Exell JC, Baxter NJ, Waltho JP & Grasby JA (2015) The Catalytic Cycle of hFEN1 Requires Protein and DNA Conformational Changes, but Are They Rate-Limiting?. PROTEIN SCIENCE, Vol. 24 (pp 147-147)
  • Waltho J (2013) Kinases, phosphatases, mutases and G-proteins. FEBS JOURNAL, Vol. 280 (pp 93-93)
  • Leigh KN, Waltho JP, Blackburn GM, Bowler MW & Webster CE (2013) Computational studies of intermediate- and transition-state analogs in b-PGM. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, Vol. 245
  • Leigh KN, Waltho JP, Blackburn GM, Bowler MW & Webster CE (2012) Utilization of computed F-19 NMR to identify active site intermediate and transition-state analogs in beta-PGM. ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, Vol. 244
  • Briggs DC, Ali T, Tongsoongnoen W, Rugg MS, Waltho JP, Levoli E, Mikecz K, Salustri A, Milner CS & Day AJ (2010) Towards the molecular basis of cumulus matrix formation: structure/function studies on TSG-6. INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, Vol. 91(6) (pp A31-A31)
  • Skerget K, Vilfan A, Waltho JP, Turk D & Zerovnik E (2008) A model for amyloid fibril formation by human stefin B (cystatin B). FEBS JOURNAL, Vol. 275 (pp 199-199)
  • WRIGHT PE, DYSON HJ, FEHER VA, TENNANT LL, WALTHO JP, LERNER RA & CASE DA (1990) FOLDING OF PEPTIDE-FRAGMENTS OF PROTEINS IN AQUEOUS-SOLUTION. FRONTIERS OF NMR IN MOLECULAR BIOLOGY, Vol. 109 (pp 1-13)
  • WRIGHT PE, DYSON HJ, WALTHO JP & LERNER RA (1990) FOLDING OF PEPTIDE-FRAGMENTS OF PROTEINS IN WATER SOLUTION. PROTEIN FOLDING : DECIPHERING THE SECOND HALF OF THE GENETIC CODE (pp 95-+)