Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase

Ming Yue Lee, James Geiger, Andrii Ishchenko, Gye Won Han, Anton Barty, Thomas A. White, Cornelius Gati, Alexander Batyuk, Mark S. Hunter, Andrew Aquila, Sébastien Boutet, Uwe Weierstall, Vadim Cherezovb, Wei Liua

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Serial femtosecond crystallography (SFX) with X-ray free-electron lasers (XFELs) has proven highly successful for structure determination of challenging membrane proteins crystallized in lipidic cubic phase; however, like most techniques, it has limitations. Here we attempt to address some of these limitations related to the use of a vacuum chamber and the need for attenuation of the XFEL beam, in order to further improve the efficiency of this method. Using an optimized SFX experimental setup in a helium atmosphere, the room-temperature structure of the adenosine A2A receptor (A2AAR) at 2.0 Å resolution is determined and compared with previous A2AAR structures determined in vacuum and/or at cryogenic temperatures. Specifically, the capability of utilizing high XFEL beam transmissions is demonstrated, in conjunction with a high dynamic range detector, to collect high-resolution SFX data while reducing crystalline material consumption and shortening the collection time required for a complete dataset. The experimental setup presented herein can be applied to future SFX applications for protein nanocrystal samples to aid in structure-based discovery efforts of therapeutic targets that are difficult to crystallize.

Original languageEnglish (US)
Pages (from-to)976-984
Number of pages9
JournalIUCrJ
Volume7
DOIs
StatePublished - Nov 1 2020

Keywords

  • G-protein-coupled receptors
  • XFELs
  • adenosine Areceptors
  • high dynamic range detectors
  • lipidic cubic phases
  • membrane proteins
  • serial femtosecond crystallography

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

Fingerprint Dive into the research topics of 'Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase'. Together they form a unique fingerprint.

Cite this