title: Serial femtosecond crystallography of soluble proteins in lipidic cubic phase.
abstract: 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.
authors: 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.
authors: Raimund
authors: R
authors: Fromme
authors: Bridge Institute, Department of Chemistry, University of Southern California , 3430 South Vermont Avenue, MC 3303, Los Angeles, CA 90089, USA.
authors: Andrii
authors: A
authors: Ishchenko
authors: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany ; Department of Physics, University of Hamburg , Luruper Chaussee 149, 22761 Hamburg, Germany.
authors: Markus
authors: M
authors: Metz
authors: 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.
authors: Shatabdi Roy
authors: SR
authors: Chowdhury
authors: 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.
authors: Shibom
authors: S
authors: Basu
authors: Linac Coherent Light Source, SLAC National Accelerator Laboratory , 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors: Sébastien
authors: S
authors: Boutet
authors: 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.
authors: Petra
authors: P
authors: Fromme
authors: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany.
authors: Thomas A
authors: TA
authors: White
authors: Center for Free Electron Laser Science, Deutsches Elektronen-Synchrotron (DESY) , Notkestrasse 85, 22607 Hamburg, Germany.
authors: Anton
authors: A
authors: Barty
authors: Department of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287, USA.
authors: John C H
authors: JC
authors: Spence
authors: Department of Physics, Arizona State University , PO Box 871504, Tempe, AZ 85287, USA.
authors: Uwe
authors: U
authors: Weierstall
authors: 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.
authors: Wei
authors: W
authors: Liu
authors: Bridge Institute, Department of Chemistry, University of Southern California , 3430 South Vermont Avenue, MC 3303, Los Angeles, CA 90089, USA.
authors: Vadim
authors: V
authors: Cherezov
keywords: X-ray free-electron laser; lipidic cubic phase; serial femtosecond crystallography; soluble protein