Development of hybrid structural-MS to determine the 3D structures of protein complexes

Hydrogen deuterium exchange MS (HDX-MS) permits the real-time characterisation of protein dynamics and solvent accessibility on the peptide level. Recent advances in HDX-MS have turned this complex procedure into a robust, high-throughput and information rich tool. The aim of this studentship is to develop ways to extract information embedded in HDX patters for use with structural elucidation and dynamical characterisation of protein complexes. The student will develop ways to characterise native state dynamics from atomic structures that will then be used as a reference point to explain non-canonical protein motions involved in function and also to elucidate structural features such as binding interfaces. HDX-MS will also be used to interrogate the folding of protein complexes such as the integrity of binding interfaces and protein folds under mildly denaturing conditions that are typically used to generate sub-complexes. Where possible electron transfer dissociation will be used to access residue-specific information and to assess the utility of protection values in HDX-MS to map the free energy surfaces of proteins and their complexes.

Labile protein protons exchange for bulk by rates determined by the chemical exchange factors and secondary structural effects. When the surrounding media is deuterium oxide these exchange events are coupled to a change in mass that can be tracked using high resolution mass spectrometry. HDXMS is important as is has the potential to allow the determination of the energy surface of a protein complex which is currently intractable by other methods. The student will be using HDXMS, high resolution MS, ion mobility mass spectrometry, native MS and potentially electron transfer dissociation. The project will have impact for Waters Corporation who design and Manufacture the instruments.