Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

Raimund Fromme; Andrii Ishchenko; Markus Metz; Shatabdi Roy Chowdhury; Shibom Basu; Sébastien Boutet; Petra Fromme; Thomas A. White; Anton Barty; John C. H. Spence; Uwe Weierstall; Wei Liu; Vadim Cherezov

doi:10.1107/S2052252515013160

IUCrJ (Sep 2015)

Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

Raimund Fromme,
Andrii Ishchenko,
Markus Metz,
Shatabdi Roy Chowdhury,
Shibom Basu,
Sébastien Boutet,
Petra Fromme,
Thomas A. White,
Anton Barty,
John C. H. Spence,
Uwe Weierstall,
Wei Liu,
Vadim Cherezov

Affiliations

Raimund Fromme: Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA
Andrii Ishchenko: Bridge Institute, Department of Chemistry, University of Southern California, 3430 South Vermont Avenue, MC 3303, Los Angeles, CA 90089, USA
Markus Metz: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany
Shatabdi Roy Chowdhury: Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA
Shibom Basu: Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA
Sébastien Boutet: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA
Petra Fromme: Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA
Thomas A. White: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany
Anton Barty: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany
John C. H. Spence: Department of Physics, Arizona State University, PO Box 871504, Tempe, AZ 85287, USA
Uwe Weierstall: Department of Physics, Arizona State University, PO Box 871504, Tempe, AZ 85287, USA
Wei Liu: Center for Applied Structural Discovery at the Biodesign Institute, Department of Chemistry and Biochemistry, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA
Vadim Cherezov: Bridge Institute, Department of Chemistry, University of Southern California, 3430 South Vermont Avenue, MC 3303, Los Angeles, CA 90089, USA

DOI: https://doi.org/10.1107/S2052252515013160
Journal volume & issue: Vol. 2, no. 5
pp. 545 – 551

Abstract

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Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP–SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP–SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.

Published in IUCrJ

ISSN: 2052-2525 (Online)
Publisher: International Union of Crystallography
Country of publisher: United Kingdom
LCC subjects: Science: Chemistry: Crystallography
Website: https://journals.iucr.org/m/

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