Serial femtosecond crystallography of G protein-coupled receptors

  • Science. 2013 Dec 20;342(6165):1521-4. doi: 10.1126/science.1244142.
Wei Liu  1 ,  Daniel Wacker ,  Cornelius Gati ,  Gye Won Han ,  Daniel James ,  Dingjie Wang ,  Garrett Nelson ,  Uwe Weierstall ,  Vsevolod Katritch ,  Anton Barty ,  Nadia A Zatsepin ,  Dianfan Li ,  Marc Messerschmidt ,  Sébastien Boutet ,  Garth J Williams ,  Jason E Koglin ,  M Marvin Seibert ,  Chong Wang ,  Syed T A Shah ,  Shibom Basu ,  Raimund Fromme ,  Christopher Kupitz ,  Kimberley N Rendek ,  Ingo Grotjohann ,  Petra Fromme ,  Richard A Kirian ,  Kenneth R Beyerlein ,  Thomas A White ,  Henry N Chapman ,  Martin Caffrey ,  John C H Spence ,  Raymond C Stevens ,  Vadim Cherezov
Affiliations
  • 1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
PMID: 24357322 DOI: 10.1126/science.1244142
Abstract

X-ray crystallography of G protein-coupled receptors and other membrane proteins is hampered by difficulties associated with growing sufficiently large crystals that withstand radiation damage and yield high-resolution data at synchrotron sources. We used an x-ray free-electron laser (XFEL) with individual 50-femtosecond-duration x-ray pulses to minimize radiation damage and obtained a high-resolution room-temperature structure of a human serotonin receptor using sub-10-micrometer microcrystals grown in a membrane mimetic matrix known as lipidic cubic phase. Compared with the structure solved by using traditional microcrystallography from cryo-cooled crystals of about two orders of magnitude larger volume, the room-temperature XFEL structure displays a distinct distribution of thermal motions and conformations of residues that likely more accurately represent the receptor structure and dynamics in a cellular environment.