Determination of structure of integral membrane proteins, especially in their native environment, is a formidable challenge in structural biology. Here we demonstrate that magic angle spinning solid-state NMR spectroscopy can be used to determine structures of membrane proteins reconstituted in synthetic lipids, an environment similar to the natural membrane. We combined a large number of experimentally determined interatomic distances and local torsional restraints to solve the structure of an oligomeric membrane protein of common seven-helical fold, Anabaena sensory rhodopsin (ASR). We determined the atomic resolution detail of the oligomerization interface of the ASR trimer, and the arrangement of helices, side chains and the retinal cofactor in the monomer.
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This research was funded by Natural Science and Engineering Research Council of Canada, National Research Foundation of Korea (Global Research Network Program), Canada Foundation for Innovation and Ontario Ministry of Economic Development and Innovation. S.W. is a recipient of the Canadian Institutes for Health Research Postdoctoral Fellowship. V.L. is supported by Canada Research Chair in Biophysics (Tier II). We thank B. Bardiaux (Leibniz-Institut für Molekulare Pharmakologie, Berlin) for providing the ARIA 2.3 program before publication, and C.P. Jaroniec and J. Lanyi for carefully reading the manuscript.
The authors declare no competing financial interests.
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Wang, S., Munro, R., Shi, L. et al. Solid-state NMR spectroscopy structure determination of a lipid-embedded heptahelical membrane protein. Nat Methods 10, 1007–1012 (2013). https://doi.org/10.1038/nmeth.2635
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