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Single particle cryo‐EM studies of membrane proteins
Author(s) -
Cheng Yifan
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.89.4
Subject(s) - lipid bilayer , nanodisc , membrane protein , integral membrane protein , chemistry , biophysics , protein–lipid interaction , ion channel , membrane , nanotechnology , biochemistry , biology , materials science , receptor
In the last few years, major technological advancements, particularly the development of new direct electron detection cameras and associated technologies, have enabled single particle electron cryo‐microscopy (cryo‐EM) to become the technique of choice for high‐resolution structure determination of many challenging biological macromolecules. We pioneered some of the technologies now being routinely used, and applied them to determine atomic structures of a number of integral membrane proteins, such as some major members of transient receptor potential (TRP) channel superfamily. Some of our studies paved the way of applying this method to a wide range of integral membrane proteins. Nowadays, atomic structures of membrane proteins that are refractory to crystallization are being determined by this method in a rapid pace. Technologically, we further aimed to enable high‐resolution structural studies of membrane proteins in near native lipid bilayer environment. To this end, we pioneered a number of approaches, including reconstituting membrane protein into lipid nanodisc or into saposin based lipid nanoparticles, and developed methodology of using conformational specific monoclonal Fab or other designed scaffoldings to facilitate image alignment of very small integral membrane proteins reconstituted into lipid nanoparticles. In a number of recent studies, we observed specific lipid‐protein interactions that play critical role in regulating the channel functions. In TRPV1 ion channel, as an example, certain specific lipid‐protein interactions enhance binding of a spider toxin to the channel through formation of a toxin‐lipid‐TRPV1 tripartite complex, and an endogenous phosphatidylinositol lipid occupies the capsaicin‐binding site of the channel. These observations provide important clues about physiological mechanisms of channel regulation. Support or Funding Information This work is funded by NIH grants: R01GM098672, S10OD020054 and S10OD021741. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .