@article{ca7d93dab54f439c85a5d9e6d19459a5,
title = "Structure of an endosomal signaling GPCR–G protein–β-arrestin megacomplex",
abstract = "Classically, G-protein-coupled receptors (GPCRs) are thought to activate G protein from the plasma membrane and are subsequently desensitized by β-arrestin (β-arr). However, some GPCRs continue to signal through G protein from internalized compartments, mediated by a GPCR–G protein–β-arr {\textquoteleft}megaplex{\textquoteright}. Nevertheless, the molecular architecture of the megaplex remains unknown. Here, we present its cryo-electron microscopy structure, which shows simultaneous engagement of human G protein and bovine β-arr to the core and phosphorylated tail, respectively, of a single active human chimeric β2-adrenergic receptor with the C-terminal tail of the arginine vasopressin type 2 receptor (β2V2R). All three components adopt their canonical active conformations, suggesting that a single megaplex GPCR is capable of simultaneously activating G protein and β-arr. Our findings provide a structural basis for GPCR-mediated sustained internalized G protein signaling.",
author = "Nguyen, {Anthony H.} and Thomsen, {Alex R.B.} and Cahill, {Thomas J.} and Rick Huang and Huang, {Li Yin} and Tara Marcink and Clarke, {Oliver B.} and S{\o}ren Heissel and Ali Masoudi and Danya Ben-Hail and Fadi Samaan and Dandey, {Venkata P.} and Tan, {Yong Zi} and Chuan Hong and Mahoney, {Jacob P.} and Sarah Triest and John Little and Xin Chen and Roger Sunahara and Jan Steyaert and Henrik Molina and Zhiheng Yu and {des Georges}, Amedee and Lefkowitz, {Robert J.}",
note = "Funding Information: We thank Q. Lennon, J. Bisson and J. Taylor for excellent administrative support, W. Capel for technical support, Y. Zhang and G. Skiniotis for help with initial sample screening, C.-R. Liang, L.-L. Gu and J.-M. Shan for synthesizing BI-167107, T. Wang and the CUNY Advanced Science Research Center Imaging Facility for help with sample screening and data collection, the lab of K. Gardner at the CUNY Advanced Science Research Center for providing various general lab reagents and equipment during initial sample screening, M. Walters, M. DeLong, M. Plue, T. Milledge, D. Capel and X. Jiang at Duke University for technical support and discussion, D. Lyumkis, D. Tegunov, W. Rice, E. Eng, L. Kim, M. Kopylov and A. Cheng for help with tilted data collection and processing and S. Houston and B. Plouffe for helpful discussion. This work received support from NIH grants (nos. T32GM007171 and F30HL149213 to A.H.N; F30HL129803 to T.J.C.; T32GM007767 to J.P.M.; R35GM133598 to A.d.G. and R01HL016037 to R.J.L.); HHMI Medical Research Fellowship to A.H.N.; the Danish Council for Independent Research & Lundbeck Foundation (DFF-5053-00136 and R172-2014-1468 to A.R.B.T.); American Heart Association Innovative Project Award (no. 19IPLOI34760706 to A.d.G); Institut de Recherche Servier (no. 18021932 to A.d.G. and D.B.-H.); and American Heart Association Predoctoral Fellowship (no. 13PRE17110027 to J.P.M.). Some of this work was performed at the Simons Electron Microscopy Center and National Resource for Automated Molecular Microscopy located at the New York Structural Biology Center, supported by grants from the Simons Foundation (grant no. SF349247), NYSTAR, and the NIH National Institute of General Medical Sciences (grant no. GM103310) with additional support from Agouron Institute (grant no. F00316) and the NIH (grant no. OD019994). R.J.L. is an HHMI Investigator. Publisher Copyright: {\textcopyright} 2019, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2019",
month = dec,
day = "1",
doi = "10.1038/s41594-019-0330-y",
language = "English (US)",
volume = "26",
pages = "1123--1131",
journal = "Nature Structural Biology",
issn = "1545-9993",
publisher = "Nature Publishing Group",
number = "12",
}