Conformation and dynamics of recombinant membrane proteins in a bacterial environment using (DNP-enhanced) cellular solid-state NMR spectroscopy

Orateur : Marie RENAULT (Institut de Pharmacologie et de Biologie Structurale/ RMN et interactions protéines-membranes, Toulouse)

Decrypting the structure, function, and molecular interactions of membrane proteins in their cellular context and at atomic resolution is of prime importance for understanding fundamental physiological processes. Solid-state NMR spectroscopy (ssNMR) is a method of choice to investigate such heterogeneous samples and has made significant progress in determining three-dimensional protein structures under in-vitro conditions. The potential of ssNMR to provide such information in-situ (i.e. cellular environment) has been revealed recently and opens new opportunities to study membrane protein structures and processes with direct implications for understanding cellular functioning and disease. Recently, we obtained structural information of a recombinant integral membrane protein and the major endogenous molecular components in a bacterial environment by combining dedicated protein production and labelling schemes with tailored ssNMR pulse methods and state-ofthe- art NMR signal enhancement procedures such as Dynamic Nuclear Polarization (DNP). Our approach allows studying entire cellular compartments as well as cell-associated proteins at the same time and at atomic resolution (in M. Baldus’s group, Utrecht University). Future challenge is to apply and develop such emerging (DNP-enhanced) ssNMR protocols on bacteria from Corynebacteriales suborder to infer the 3D structure and molecular interactions of membrane proteins of high pharmacological interest in their native (cellular) environment (in A. Milon’s group, IPBS UMR-CNRS 5089/ Toulouse).

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