Journal article
Phase-plate cryo-EM structure of a class B GPCR-G-protein complex
YL Liang, M Khoshouei, M Radjainia, Y Zhang, A Glukhova, J Tarrasch, DM Thal, SGB Furness, G Christopoulos, T Coudrat, R Danev, W Baumeister, LJ Miller, A Christopoulos, BK Kobilka, D Wootten, G Skiniotis, PM Sexton
Nature | NATURE PUBLISHING GROUP | Published : 2017
DOI: 10.1038/nature22327
Abstract
Class B G-protein-coupled receptors are major targets for the treatment of chronic diseases, such as osteoporosis, diabetes and obesity. Here we report the structure of a full-length class B receptor, the calcitonin receptor, in complex with peptide ligand and heterotrimeric Gα s βÎ 3 protein determined by Volta phase-plate single-particle cryo-electron microscopy. The peptide agonist engages the receptor by binding to an extended hydrophobic pocket facilitated by the large outward movement of the extracellular ends of transmembrane helices 6 and 7. This conformation is accompanied by a 60° kink in helix 6 and a large outward movement of the intracellular end of this helix, opening the bundl..
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Awarded by National Institutes of Health
Funding Acknowledgements
This work was funded by the National Health and Medical Research Council of Australia (NHMRC) (grant numbers 1055134, 1061044 and 1120919) and NIH grants DK090165, NS092695. P.M.S. and A.C. are NHMRC Principal and Senior Principal Research Fellows, respectively. D.W. is a NHMRC Career Development Fellow. Computational studies were partially supported by Melbourne Bioinformatics at the University of Melbourne, grant number VR0024. Negative-stain imaging and cryo-EM screening was performed at the Monash Ramaciotti Centre for Cryo-Electron Microscopy. The GPCRdb (http://gpcrdb.org) was used for generation of initial alignments of human class B GPCR sequences. The authors thank the late M. Azria for 100 mg of sCT, used in initial work.