Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors

X Edward Zhou; Yuanzheng He; Parker W de Waal; Xiang Gao; Yanyong Kang; Ned Van Eps; Yanting Yin; Kuntal Pal; Devrishi Goswami; Thomas A White; Anton Barty; Naomi R Latorraca; Henry N Chapman; Wayne L Hubbell; Ron O Dror; Raymond C Stevens; Vadim Cherezov; Vsevolod V Gurevich; Patrick R Griffin; Oliver P Ernst; Karsten Melcher; H Eric Xu

doi:10.1016/j.cell.2017.07.002

Identification of Phosphorylation Codes for Arrestin Recruitment by G Protein-Coupled Receptors

Cell. 2017 Jul 27;170(3):457-469.e13. doi: 10.1016/j.cell.2017.07.002.

Authors

X Edward Zhou¹, Yuanzheng He², Parker W de Waal², Xiang Gao², Yanyong Kang², Ned Van Eps³, Yanting Yin¹, Kuntal Pal², Devrishi Goswami⁴, Thomas A White⁵, Anton Barty⁵, Naomi R Latorraca⁶, Henry N Chapman⁷, Wayne L Hubbell⁸, Ron O Dror⁶, Raymond C Stevens⁹, Vadim Cherezov¹⁰, Vsevolod V Gurevich¹¹, Patrick R Griffin⁴, Oliver P Ernst¹², Karsten Melcher², H Eric Xu¹³

Affiliations

¹ VARI-SIMM Center, Center for Structure and Function of Drug Targets, CAS-Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, MI 49503, USA.
² Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, MI 49503, USA.
³ Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada.
⁴ Department of Molecular Medicine, The Scripps Research Institute, Scripps Florida, Jupiter, FL 33458, USA.
⁵ Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
⁶ Department of Computer Science, Stanford University, Stanford, CA 94305, USA; Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA; Department of Structural Biology, Stanford University, Stanford, CA 94305, USA; Institute for Computational and Mathematical Engineering, Stanford University, Stanford, CA 94305, USA; Biophysics Program, Stanford University, Stanford, CA 94305, USA.
⁷ Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany; Centre for Ultrafast Imaging, 22761 Hamburg, Germany.
⁸ Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA.
⁹ Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA; iHuman Institute, ShanghaiTech University, 2F Building 6, 99 Haike Road, Pudong New District, Shanghai 201210, China.
¹⁰ Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, CA 90089, USA.
¹¹ Department of Pharmacology, Vanderbilt University, Nashville, TN 37232, USA.
¹² Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
¹³ VARI-SIMM Center, Center for Structure and Function of Drug Targets, CAS-Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, MI 49503, USA. Electronic address: eric.xu@vai.org.

Abstract

G protein-coupled receptors (GPCRs) mediate diverse signaling in part through interaction with arrestins, whose binding promotes receptor internalization and signaling through G protein-independent pathways. High-affinity arrestin binding requires receptor phosphorylation, often at the receptor's C-terminal tail. Here, we report an X-ray free electron laser (XFEL) crystal structure of the rhodopsin-arrestin complex, in which the phosphorylated C terminus of rhodopsin forms an extended intermolecular β sheet with the N-terminal β strands of arrestin. Phosphorylation was detected at rhodopsin C-terminal tail residues T336 and S338. These two phospho-residues, together with E341, form an extensive network of electrostatic interactions with three positively charged pockets in arrestin in a mode that resembles binding of the phosphorylated vasopressin-2 receptor tail to β-arrestin-1. Based on these observations, we derived and validated a set of phosphorylation codes that serve as a common mechanism for phosphorylation-dependent recruitment of arrestins by GPCRs.

Keywords: GPCR; GRK; arrestin; biased signaling; drug discovery; membrane proteins; phosphorylation codes; rhodopsin.

MeSH terms

Amino Acid Sequence
Animals
Arrestins / chemistry*
Arrestins / metabolism
Chromatography, Liquid
Humans
Mice
Models, Molecular
Phosphorylation
Rats
Rhodopsin / chemistry*
Rhodopsin / metabolism
Sequence Alignment
Tandem Mass Spectrometry
X-Rays

Substances

Arrestins
arrestin 1 protein, mouse
Rhodopsin

Abstract

MeSH terms

Substances

Grants and funding