1. Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase
- Author
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Alexander Batyuk, Gye Won Han, Andrii Ishchenko, Mark S. Hunter, Sébastien Boutet, Vadim Cherezov, Andrew Aquila, Uwe Weierstall, Cornelius Gati, Wei Liu, Anton Barty, Thomas A. White, James Geiger, and Ming-Yue Lee
- Subjects
0301 basic medicine ,serial femtosecond crystallography ,Materials science ,g-protein-coupled receptors ,membrane proteins ,02 engineering and technology ,Biochemistry ,law.invention ,X-ray laser ,03 medical and health sciences ,adenosine a2a receptors ,law ,General Materials Science ,ddc:530 ,lcsh:Science ,Dynamic range ,lipidic cubic phases ,Resolution (electron density) ,Detector ,General Chemistry ,high dynamic range detectors ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,Laser ,xfels ,Research Letters ,Crystallography ,030104 developmental biology ,Nanocrystal ,Femtosecond ,Vacuum chamber ,lcsh:Q ,0210 nano-technology - Abstract
IUCrJ 7(6), 976 - 984 (2020). doi:10.1107/S2052252520012701, 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., Published by Chester
- Published
- 2020