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

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

doi:10.1107/S2052252520012701

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

Journal Article · Thu Oct 15 00:00:00 EDT 2020 · IUCrJ

DOI:https://doi.org/10.1107/S2052252520012701· OSTI ID:1675009

; Geiger, James;

;

; Barty, Anton; White, Thomas A.;

; Batyuk, Alexander; Hunter, Mark S.; Aquila, Andrew;

; Weierstall, Uwe;

; Liu, Wei

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 A_2A receptor (A_2AAR) at 2.0 Å resolution is determined and compared with previous A_2AAR 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.

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Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Linac Coherent Light Source (LCLS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH); National Science Foundation (NSF)

Grant/Contract Number:: AC02-76SF00515; R35 GM127086; R01 GM124152; R21 DA042298; 1231306

OSTI ID:: 1675009

Alternate ID(s):: OSTI ID: 1768894

Journal Information:: IUCrJ, Journal Name: IUCrJ Vol. 7 Journal Issue: 6; ISSN 2052-2525

Publisher:: International Union of CrystallographyCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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