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Sci Data. 2016 Apr 12;3:160021. doi: 10.1038/sdata.2016.21.

X-ray laser diffraction for structure determination of the rhodopsin-arrestin complex.

Author information

1
Laboratory of Structural Sciences, Center for Structural Biology and Drug Discovery, Van Andel Research Institute, Grand Rapids, Michigan 49503, USA.
2
Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
3
School of Molecular Sciences, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
4
Department of Chemistry, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
5
Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, California 90089, USA.
6
Joint Center for Structural Genomics, Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
7
Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
8
Swiss Light Source at Paul Scherrer Institute, CH-5232 Villigen, Switzerland.
9
School of Medicine and School of Biochemistry and Immunology, Trinity College, Dublin D02 R590, Ireland.
10
Centre for Ultrafast Imaging, 22761 Hamburg, Germany.
11
Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
12
iHuman Institute, Shanghai Tech University, 2F Building 6, 99 Haike Road, Pudong New District, Shanghai 201210, China.
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.

Abstract

Serial femtosecond X-ray crystallography (SFX) using an X-ray free electron laser (XFEL) is a recent advancement in structural biology for solving crystal structures of challenging membrane proteins, including G-protein coupled receptors (GPCRs), which often only produce microcrystals. An XFEL delivers highly intense X-ray pulses of femtosecond duration short enough to enable the collection of single diffraction images before significant radiation damage to crystals sets in. Here we report the deposition of the XFEL data and provide further details on crystallization, XFEL data collection and analysis, structure determination, and the validation of the structural model. The rhodopsin-arrestin crystal structure solved with SFX represents the first near-atomic resolution structure of a GPCR-arrestin complex, provides structural insights into understanding of arrestin-mediated GPCR signaling, and demonstrates the great potential of this SFX-XFEL technology for accelerating crystal structure determination of challenging proteins and protein complexes.

PMID:
27070998
PMCID:
PMC4828943
DOI:
10.1038/sdata.2016.21
[Indexed for MEDLINE]
Free PMC Article

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