Probing membrane protein ground and conformationally excited states using dipolar- and J-coupling mediated MAS solid state NMR experiments

Methods : a Companion to Methods in Enzymology
T Gopinath, Gianluigi Veglia

Abstract

The intrinsic conformational plasticity of membrane proteins directly influences the magnitude of the orientational-dependent NMR interactions such as dipolar couplings (DC) and chemical shift anisotropy (CSA). As a result, the conventional cross-polarization (CP)-based techniques mainly capture the more rigid regions of membrane proteins, while the most dynamic regions are essentially invisible. Nonetheless, dynamic regions can be detected using experiments in which polarization transfer takes place via J-coupling interactions. Here, we review our recent efforts to develop single and dual acquisition pulse sequences with either 1H or 13C detection that utilize both DC and J-coupling mediated transfer to detect both rigid and mobile regions of membrane proteins in native-like lipid environments. We show the application of these new methods for studying the conformational equilibrium of a single-pass membrane protein, phospholamban, which regulates the calcium transport across the sarcoplasmic reticulum (SR) membrane by interacting with the SR Ca2+-ATPase. We anticipate that these methods will be ideal to portray the complex dynamics of membrane proteins in their native environments.

Citations

Jul 22, 2020·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Martin DračínskýPaul Hodgkinson
Aug 11, 2019·Biochimica Et Biophysica Acta. Biomembranes·Erik K LarsenGianluigi Veglia
Mar 4, 2021·Accounts of Chemical Research·Tata GopinathGianluigi Veglia
Aug 17, 2021·Biochemical Society Transactions·Vivien Yeh, Boyan B Bonev

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