title: Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser.
abstract: G-protein-coupled receptors (GPCRs) signal primarily through G proteins or arrestins. Arrestin binding to GPCRs blocks G protein interaction and redirects signalling to numerous G-protein-independent pathways. Here we report the crystal structure of a constitutively active form of human rhodopsin bound to a pre-activated form of the mouse visual arrestin, determined by serial femtosecond X-ray laser crystallography. Together with extensive biochemical and mutagenesis data, the structure reveals an overall architecture of the rhodopsin-arrestin assembly in which rhodopsin uses distinct structural elements, including transmembrane helix 7 and helix 8, to recruit arrestin. Correspondingly, arrestin adopts the pre-activated conformation, with a ∼20° rotation between the amino and carboxy domains, which opens up a cleft in arrestin to accommodate a short helix formed by the second intracellular loop of rhodopsin. This structure provides a basis for understanding GPCR-mediated arrestin-biased signalling and demonstrates the power of X-ray lasers for advancing the frontiers of structural biology.
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Yanyong
authors.initials: Y
authors.lastname: Kang
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: X Edward
authors.initials: XE
authors.lastname: Zhou
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Yuanzheng
authors.initials: Y
authors.lastname: He
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Andrii
authors.initials: A
authors.lastname: Ishchenko
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: White
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Yefanov
authors.affiliation: Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Gye Won
authors.initials: GW
authors.lastname: Han
authors.affiliation: Joint Center for Structural Genomics, Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Qingping
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Parker W
authors.initials: PW
authors.lastname: de Waal
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Jiyuan
authors.initials: J
authors.lastname: Ke
authors.affiliation: 1] Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA [2] Department of Obstetrics &Gynecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: M H Eileen
authors.initials: MH
authors.lastname: Tan
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Chenghai
authors.initials: C
authors.lastname: Zhang
authors.affiliation: The National Resource for Automated Molecular Microscopy, New York Structural Biology Center, New York, New York 10027, USA.
authors.forename: Arne
authors.initials: A
authors.lastname: Moeller
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA.
authors.forename: Graham M
authors.initials: GM
authors.lastname: West
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA.
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Pascal
authors.affiliation: Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Ned
authors.initials: N
authors.lastname: Van Eps
authors.affiliation: Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Lydia N
authors.initials: LN
authors.lastname: Caro
authors.affiliation: Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232, USA.
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Vishnivetskiy
authors.affiliation: Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232, USA.
authors.forename: Regina J
authors.initials: RJ
authors.lastname: Lee
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Suino-Powell
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Gu
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Kuntal
authors.initials: K
authors.lastname: Pal
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Jinming
authors.initials: J
authors.lastname: Ma
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Zhi
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: 1] Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA [2] BioXFEL, NSF Science and Technology Center, 700 Ellicott Street, Buffalo, New York 14203, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Messerschmidt
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Cornelius
authors.initials: C
authors.lastname: Gati
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Nadia A
authors.initials: NA
authors.lastname: Zatsepin
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Dingjie
authors.initials: D
authors.lastname: Wang
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: James
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Shibom
authors.initials: S
authors.lastname: Basu
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Shatabdi
authors.initials: S
authors.lastname: Roy-Chowdhury
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Chelsie E
authors.initials: CE
authors.lastname: Conrad
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Coe
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Beijing Computational Science Research Center, Haidian District, Beijing 10084, China.
authors.forename: Haiguang
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Stella
authors.initials: S
authors.lastname: Lisova
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53211, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Kupitz
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Ingo
authors.initials: I
authors.lastname: Grotjohann
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Raimund
authors.initials: R
authors.lastname: Fromme
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Minjia
authors.initials: M
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Obstetrics &Gynecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Swiss Light Source at Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Meitian
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Zheng
authors.affiliation: School of Medicine and School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland.
authors.forename: Dianfan
authors.initials: D
authors.lastname: Li
authors.affiliation: School of Medicine and School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland.
authors.forename: Nicole
authors.initials: N
authors.lastname: Howe
authors.affiliation: 1] BioXFEL, NSF Science and Technology Center, 700 Ellicott Street, Buffalo, New York 14203, USA [2] Ben May Department for Cancer Research, University of Chicago, Chicago, Illinois 60637, USA.
authors.forename: Yingming
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Laboratory of Biomolecular Research at Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
authors.forename: Jörg
authors.initials: J
authors.lastname: Standfuss
authors.affiliation: Department of Biology, Universität Konstanz, 78457 Konstanz, Germany.
authors.forename: Kay
authors.initials: K
authors.lastname: Diederichs
authors.affiliation: Beijing Synchrotron Radiation Facility, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Dong
authors.affiliation: The National Resource for Automated Molecular Microscopy, New York Structural Biology Center, New York, New York 10027, USA.
authors.forename: Clinton S
authors.initials: CS
authors.lastname: Potter
authors.affiliation: The National Resource for Automated Molecular Microscopy, New York Structural Biology Center, New York, New York 10027, USA.
authors.forename: Bridget
authors.initials: B
authors.lastname: Carragher
authors.affiliation: School of Medicine and School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland.
authors.forename: Martin
authors.initials: M
authors.lastname: Caffrey
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Hualiang
authors.initials: H
authors.lastname: Jiang
authors.affiliation: 1] Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany [2] Centre for Ultrafast Imaging, 22761 Hamburg, Germany.
authors.forename: Henry N
authors.initials: HN
authors.lastname: Chapman
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: John C H
authors.initials: JC
authors.lastname: Spence
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Petra
authors.initials: P
authors.lastname: Fromme
authors.affiliation: 1] Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA [2] Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Uwe
authors.initials: U
authors.lastname: Weierstall
authors.affiliation: 1] Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada [2] Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: Oliver P
authors.initials: OP
authors.lastname: Ernst
authors.affiliation: Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Vsevolod
authors.initials: V
authors.lastname: Katritch
authors.affiliation: Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232, USA.
authors.forename: Vsevolod V
authors.initials: VV
authors.lastname: Gurevich
authors.affiliation: Department of Molecular Therapeutics, The Scripps Research Institute, Scripps Florida, Jupiter, Florida 33458, USA.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Griffin
authors.affiliation: Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095, USA.
authors.forename: Wayne L
authors.initials: WL
authors.lastname: Hubbell
authors.affiliation: 1] Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA [2] Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA [3] iHuman Institute, ShanghaiTech University, 2F Building 6, 99 Haike Road, Pudong New District, Shanghai 201210, China.
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Stevens
authors.affiliation: Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Vadim
authors.initials: V
authors.lastname: Cherezov
authors.affiliation: Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
authors.forename: Karsten
authors.initials: K
authors.lastname: Melcher
authors.affiliation: 1] Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA [2] 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.
authors.forename: H Eric
authors.initials: HE
authors.lastname: Xu
mesh_terms: D000818:Animals; D019393:Arrestin / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007834:Lasers; D051379:Mice; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D015203:Reproducibility of Results; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism*; D015398:Signal Transduction; D014965:X-Rays
chemical_list: D019393:Arrestin; D004220:Disulfides; D046912:Multiprotein Complexes; D012243:Rhodopsin