Next-generation mass spectrometry of protein structure and interactions

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A major challenge at the interface of physical and life science is to understand how the function of proteins and other biomolecules is encoded in their structures and dynamics. These biomolecules are built from a small number of building blocks (such as amino acids and monosaccharides), which combine in a multitude of ways to create the vast diversity of interacting components that orchestrate processes necessary for life and responsible for malfunction in disease. A valuable tool for in-depth investigation of biological processes is mass spectrometry (MS), which provides fundamental information about the mass, identity and structure of biomolecules. With applications across almost all branches of science, mass spectrometers enable research into the mechanisms of health and disease, as well as drug discovery.To upgrade the capabilities of the advanced MS centre of excellence in the University of Oxford's Department of Chemistry, we seek to purchase a state-of-the-art mass spectrometer unrivalled in its ability to perform multidimensional analysis of protein stoichiometry, structure and dynamics. The instrument integrates MS with another high-resolution characterisation method, ion mobility spectrometry (IMS). IMS enables the measurement of molecular size based on the time taken to traverse a field of gas. In the cyclic implementation (cyclic IMS) we seek to obtain, ions can be passed indefinitely around a "race track", which dramatically increases the measurement resolution. The instrument will offer complementary capabilities (cyclic IMS, tandem MS, collision induced dissociation, electron capture dissociation and hydrogen deuterium exchange), which can be combined to tackle a variety of complex analytical problems. With cyclic IMS, for example, ions can be extracted from the ring, fragmented and then reinjected for further measurement. This capability makes it possible to probe the conformation-specific interactions of assemblies of macromolecules, revealing how subtle differences in structure correspond to differences in function.The first of its kind in the world, this instrument will bring a step change in our ability to interrogate complex biomolecular assemblies. It can perform all the measurements possible on our existing instruments, but at ten-fold higher resolution, and enable completely different types of experiments. This multi-user instrument will significantly boost the quality and capacity of research at the world-leading centre for gas-phase biophysics and structural biology. It will allow researchers within Oxford (across multiple departments and divisions) and the wider molecular characterisation community to capitalise on the latest developments in MS - to obtain measurements at unparalleled resolution, and probe the conformation-specific interactions of assemblies of proteins and other macromolecules at an unprecedented level of detail. These capabilities will make it possible to gain completely new insights into the mechanisms of function of macromolecules that play essential roles in health and disease, and often represent valuable therapeutic targets.