Cryo-EM analysis of a membrane protein embedded in the liposome

Proceedings of the National Academy of Sciences of the United States of America
Xia YaoNieng Yan

Abstract

Membrane proteins (MPs) used to be the most difficult targets for structural biology when X-ray crystallography was the mainstream approach. With the resolution revolution of single-particle electron cryo-microscopy (cryo-EM), rapid progress has been made for structural elucidation of isolated MPs. The next challenge is to preserve the electrochemical gradients and membrane curvature for a comprehensive structural elucidation of MPs that rely on these chemical and physical properties for their biological functions. Toward this goal, here we present a convenient workflow for cryo-EM structural analysis of MPs embedded in liposomes, using the well-characterized AcrB as a prototype. Combining optimized proteoliposome isolation, cryo-sample preparation on graphene grids, and an efficient particle selection strategy, the three-dimensional (3D) reconstruction of AcrB embedded in liposomes was obtained at 3.9 Å resolution. The conformation of the homotrimeric AcrB remains the same when the surrounding membranes display different curvatures. Our approach, which can be widely applied to cryo-EM analysis of MPs with distinctive soluble domains, lays out the foundation for cryo-EM analysis of integral or peripheral MPs whose functions are...Continue Reading

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Oct 2, 2020·Nature Reviews. Microbiology·Matthew T K Hankins, Maike Bublitz
Oct 21, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Andrew J Y JonesDaniel Nietlispach
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Sep 16, 2021·Journal of Chemical Theory and Computation·T Bertie AnsellRobin A Corey
Oct 9, 2021·Emerging Topics in Life Sciences·James Birch, Andrew Quigley

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