High resolution cyclic ion mobility HDX mass spectrometry of protein dynamics and function

This proposal is to enable cyclic ion mobility hydrogen deuterium exchange mass spectrometry (cyclic IMS HDX-MS) at the University of Manchester. The bid aims to increase capacity and capability for high-quality HDX-MS analysis of increasingly complex and heterogeneous biological samples, largely intractable by other biophysical and biochemical methods. It will also support world-class research at the interface between cellular biology, structural chemistry and molecular medicine with direct implications into human health and disease. The instrumentation will allow us to measure the exchange of hydrogen for heavier deuterium at high resolution enabling direct insights into how proteins move and interact with other biomolecules.

Over the past few years, the PI has pioneered the study of challenging membrane proteins and their interactions with their surrounding environment using emerging HDX-MS. As such the PI is at the best place o lead this bid not only in the UK but also worldwide. The instrumentation will allow taking the research to an entirely new level by targeting complex and increasingly larger biological systems in the native environment where they live and function. This will open up directions in understanding the mechanism of a multitude of diseases outlined in the case for support.

This HDX-MS instrument will be housed at the Manchester Institute for Biotechnology (MIB) and in the Michael Barber Centre for Collaborative Mass Spectrometry. MIB is the home of research groups from both the faculty of science and engineering (FSE) and the faculty of biology, medicine and health (FBMH). The instrument will be accessible by research groups from both FSE and FBMH and by external collaborators from both academia and industry. It will essentially form a basis for an interdisciplinary network of collaborations within the University of Manchester and beyond. We will use this state-of-the-art equipment to underpin investments by MRC and enable working with industrial partners (e.g. OMass Therapeutics, Waters Corp., BioShape Ltd, AstraZeneca, FujiDiosynth). The new instrument will enhance capacity and capability for investigating challenging biological systems within their native cellular environment. It will offer unsurpassed sensitivity, speed of acquisition and resolution circumventing current limitations in the technique. This will enable broader research access and wider application to many biological processes such as membrane transport, drug development and design, catalysis, and large molecular machines and related diseases such as cancer, tuberculosis, heart failure and antimicrobial resistance. This will further offer the ability to carry out research in human samples and in their native environments.

We request a state-of-the-art cyclic ion mobility quadrupole time of flight (QTof) mass spectrometer equipped with a Trajan Robot system for HDX automation, and a lipid filtration system for automated analysis of challenging membrane proteins within their native lipid environments. The equipment will enable HDX-MS experiments at enhanced sensitivity and resolution. The first will be obtained at ~ 5-fold less sample compared to other similar instruments important for membrane proteins that often suffer by low yields of purification. The latter will be afforded by the multiple-pass ion mobility capabilities. These unique analytical capabilities together with the speed of acquisition offered by this bespoke instrumentation will allow us to timely analyse complex biological samples such as membrane proteins, protein lipid assemblies, and high-mass multi-protein complexes. Importantly, the equipment will afford us unprecedented data quality and interrogate the functional dynamics of a wide range of proteins at physiological conditions and at increased mass ranges, circumventing current limitations of HDX-MS (e.g. peak capacity). This will make inroads into better understanding diverse disease states and many pathologies.