Unravelling the structure and conformational dynamics of membrane proteins using H/D-exchange mass spectrometry and cryo-EM

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Membrane proteins are biological macromolecules that are present within dynamic and complex cellular membranes. They have diverse complex cellular functions and represent more than half of drug targets; however, the molecular mechanisms governing their modes of action remain poorly understood. Most membrane proteins are highly dynamic and undergo structural and conformational rearrangements to perform their function. Whilst Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) and Cryo-Electron Microscopy (cryo-EM) each provide unique information, both techniques complement and validate each other providing static pictures as well as dynamics and flexibility information of large and complex systems1. This proposal focuses on developing workflows to understand the structure and conformational dynamics of membrane proteins using HDX-MS and cryo-EM. This project will focus on developing the knowledge and expertise of the PhD student in both HDX-MS and cryo-EM by studying the efflux pump (MdtF). The first HDX-MS and native MS measurements of membrane proteins (including AcrB) in SMALPs2,3 have been published as well as a recent publication demonstrating the impact of a clinically relevant efflux pump mutation in AcrB using HDX-MS4.