Applications of NMR Spectroscopy to Study Structure, Dynamics and Small Molecule Interactions Related to Protein Folding and Misfolding

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NMR spectroscopy is a powerful technique to study the structure, dynamics and interactions of biomacromolecules in their natural environment in solution, including proteins the cellular milieu, and in the solid-state, including biological membranes, biomaterials and membrane-proteins. This methodology allows researchers to gain detailed atomic resolution pictures of these components and to also study the strength and frequency of the interactions incurred by these biomolecules, giving a comprehensive picture of the systems of interest. A unique feature of the solution biomolecular NMR, in particular recently developed TXO cryoprobes, is the ability to characterise residual structure and transient interactions by proteins which lack distinct three dimensional structures and which are referred to as intrinsically disordered. Intrinsic disorder is a primary characteristic of ~30% of the proteins encoded in the eukaryotic genome and these proteins are central to a number of cellular processes as well as aberrant human diseases including cancer, neurodegenerative disorders, dilated cardiomyopathies. Another key research area in biomolecular NMR is within the solid-state; where samples are in more rigid conformations due to self-assembly or interactions with biological membranes or protein networks. Biomolecular ssNMR has been driving the characterisation of protein amyloid fibrils and membrane proteins, and is an extremely accurate method to probe the structural properties of biomolecular assemblies, i.e.biomaterials, large protein complexes and hydrogels. The NMR facility in the Department of Chemistry has been serving a large community of molecular scientists in the Cambridge area, including the Departments of Chemistry, the Centre for Misfolding Disease (CMD) and a number of companies and other Departments. Recent research developments in the research labs of the applicants have enabled significant discoveries in the field of protein aggregation and misfolding diseases, for which the CMD has been funded to boost the transitional research into finding cures for neurodegenerative disorders include Alzheimer's and Parkinson's diseases. The activities of the CMD, as well as of the other research labs accessing our NMR facility, would greatly benefit from the availability of high-throughput screening of small molecules on the precursor proteins linked with these diseases. This methodology cannot, currently, be applied with efficiency, despite our flagship 700 MHz instrument being equipped with a cryoprobe TXO and sample exchanger, and we need a more modern spectrometer console to boost sensitivity and apply fast methods of data acquisitions. The change of console is part of our bigger plan to rejuvenate our 700 MHz spectrometer, including maintenance to the magnet to ensure more cost effective management of the instrument and avoid future breakage.In addition to our expertise in solution NMR, we have gained internationally relevant expertise in studying the aggregated species that are associated with Alzheimer's and Parkinson's disorders. In this context, we would like to equip our NMR facility with bio-solid NMR capability for protein investigations, which is surprisingly missing in the Cambridge area. This equipment will allow also, the drug-screening of selected molecules to specific protein aggregates associated with Alzheimer's and Parkinson's. In addition to the great benefits that this application would have on the CMD activities associated with the study of intrinsically disordered proteins and protein aggregation in neurodegenerative disorders, the new NMR capability will serve the larger community of scientists at the University of Cambridge and the collaborating institutions that frequently access this facility, including industrial collaborators, faciliting and impacting on research encompassing various aspects of organic chemistry,material sciences and chemical biology.