Research Guides

Department of Chemistry University of Oxford

Professor J.L.P. Benesch

We are interested in studying the structure and dynamics of the cellular protein machinery. Through this we hope to understand the molecular basis for their function in healthy organisms, and malfunction in disease. Our research relies on combining traditional structural biology approaches with new biophysical techniques in order to interrogate our system of interest from multiple angles. In this way we can obtain a thorough description of the molecular structure and its associated dynamics.

Protein Misfolding Disease and Molecular Chaperones

Many maladies, ranging from Alzheimer's disease to alcoholic hepatitis, are associated with improperly folded proteins, and the aggregates they form. The body typically acts to prevent or reverse such aberrant associations, primarily through the action of the 'molecular chaperones'. These molecules, themselves proteins, act in different ways, including: ensuring faithful initial folding; trapping misfolded species; or disaggregating protein deposits. One class of chaperone, the small heat shock proteins, are implicated in a number of diseases, yet remain functionally rather poorly understood. This is due primarily to these proteins being extremely dynamic and structurally variable. We are investigating their molecular details and mechanism of their chaperone action, and thereby the basis for their malfunction in disease.

Concurrent Determination of Quaternary Structure and Dynamics

Our approach is based primarily on using mass spectrometry detection, with prior solution- and gas-phase manipulation. The speed of MS detection means that a considerable amount of structural information can be obtained in real-time. One of our research interests regards developing time-resolved MS approaches for monitoring pre-equilibrium states. In order to complement these insights on the quaternary level of protein organization with information on the amino acid level we perform additional spectrometric and spectroscopic experiments in parallel. In this way we aim to directly track the evolution of interactions and structure as a biological system reacts to a given stimulus.

Manipulating Protein Assemblies in the Gas Phase

The dissociation of macromolecular assemblies by activation in the gas phase is a useful means for obtaining details as to their organization. However it is our view that the full potential of this approach remains some distance from being fully realized, primarily due to technological limitations, and an incomplete understanding of the dissociation mechanism. We are currently focusing on both these aspects in order to maximize the information accessible from such experiments.



Quaternary dynamics and plasticity underlie small heat shock protein chaperone function
Stengel, F., Baldwin, A.J., Painter, A.J., Jaya, N., Basha, E., Kay, L.E., Vierling, E., Robinson, C.V. & Benesch, J.L.P.
Proc. Natl. Acad. Sci. U.S.A. (2010), 107, 2007-12

Collisional activation of protein complexes: picking up the pieces
Benesch, J.L.P.
J. Am. Soc. Mass. Spectrom. (2009), 20, 341-8

Small heat shock protein activity is regulated by variable oligomeric substructure
Benesch, J.L.P., Ayoub, M., Robinson, C.V. & Aquilina, J.A.
J. Biol. Chem. (2008), 283, 28513-7

Real-time monitoring of protein complexes reveals their quaternary organization and dynamics
Painter, A.J., Jaya, N., Basha, E., Vierling, E., Robinson, C.V. & Benesch, J.L.P.
Chem. Biol. (2008), 15, 246-53

Protein complexes in the gas phase: technology for structural genomics and proteomics
Benesch, J.L.P., Ruotolo, B.T., Simmons, D.A. & Robinson, C.V.
Chem. Rev. (2007), 107, 3544-67