Deciphering the conformational mechanisms of nascent membrane protein folding

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Cellular membranes are dynamic structures consisting mostly of protein and lipid which act to compartmentalise the cell, providing barriers to the external environments of the cell and its organelles. Integral membrane proteins reside within a cellular membrane and are responsible for a variety of dynamic processes, such as sensation, cellular regulation, and cell-to-cell adhesion. A membrane protein's functional capability and their level of expression will largely decide the ionic composition, and therefore the metabolic levels of a given cell type, making them essential for all life. Membrane proteins must be folded 'correctly' to become functional. The key aim of the research proposal is to understand how nascent membrane proteins fold. The chemical principles of membrane protein folding during their production and in the context of a cellular membrane are poorly understood. Investigating membrane proteins is a difficult task due to the intractable and hydrophobic nature of membrane proteins, compared to their soluble protein counter parts; with additional protein machinery being required for their cellular localisation and folding within the membrane, in comparison. To understand the processes involved would enhance our understanding on how the cell obtains 'correctly' folded and functional membrane proteins, and postulate on how 'incorrectly' misfolded and aggregated membrane proteins are formed - a toxic process for a cell. Consequently, development of robust assays that reflect the cellular processes will enable the influence of co-factors, mutations and drugs on the process of membrane protein folding. Moreover, there is a key aim to develop methodologies to gain structural insight into membrane protein events using advanced chemical tools and techniques, such as hydrogen-deuterium exchange mass spectrometry.The research is proposed to take place at the King's College London, Department of Chemistry, within the laboratory of Professor Paula Booth. The research will involve producing systems capable of both membrane protein production, insertion and folding in vitro and developing methodologies for the structural assessment of these processes with strong focus on using hydrogen deuterium exchange mass spectrometry.