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

By Ming-Yue Lee, James Geiger, Andrii Ishchenko, Gye Won Han, Anton Barty, Thomas A. White, Cornelius Gati, Alexander Batyuk, Mark S. Hunter1, Andrew Aquila, Sébastien Boutet, Uwe Weierstall2, Vadim Cherezov3, Wei Liu2

1. SLAC National Accelerator Laboratory 2. Arizona State University 3. Bridge Institute - University of Southern California

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Type

journal-article

Author

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

Citation

Lee, M.-Y. et al., 2020. Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase. IUCrJ, 7(6), pp.976–984. Available at: http://dx.doi.org/10.1107/s2052252520012701.

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.

DOI

Funding

NSF-STC Biology with X-ray Lasers (NSF-1231306)