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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 shown 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:
 [1];  [2];  [3];  [1];  [1];  [4];  [1];  [5];  [5];  [6];  [6];  [1];  [2]
  1. Arizona State Univ., Tempe, AZ (United States)
  2. Univ. of Southern California, Los Angeles, CA (United States)
  3. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  4. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  5. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  6. Arizona State Univ., Mesa, AZ (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1224055
Grant/Contract Number:  
AC03-76SF00515
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 2; Journal Issue: 5; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English
Subject:
43 PARTICLE ACCELERATORS; serial femtosecond crystallography; X-ray free-electron laser; lipidic cubic phase; soluble protein

Citation Formats

Fromme, Raimund, Ishchenko, Andrii, Metz, Markus, Chowdhury, Shatabdi Roy, Basu, Shibom, Boutet, Sébastien, Fromme, Petra, White, Thomas A., Barty, Anton, Spence, John C. H., Weierstall, Uwe, Liu, Wei, and Cherezov, Vadim. Serial femtosecond crystallography of soluble proteins in lipidic cubic phase. United States: N. p., 2015. Web. doi:10.1107/S2052252515013160.
Fromme, Raimund, Ishchenko, Andrii, Metz, Markus, Chowdhury, Shatabdi Roy, Basu, Shibom, Boutet, Sébastien, Fromme, Petra, White, Thomas A., Barty, Anton, Spence, John C. H., Weierstall, Uwe, Liu, Wei, & Cherezov, Vadim. Serial femtosecond crystallography of soluble proteins in lipidic cubic phase. United States. doi:10.1107/S2052252515013160.
Fromme, Raimund, Ishchenko, Andrii, Metz, Markus, Chowdhury, Shatabdi Roy, Basu, Shibom, Boutet, Sébastien, Fromme, Petra, White, Thomas A., Barty, Anton, Spence, John C. H., Weierstall, Uwe, Liu, Wei, and Cherezov, Vadim. Tue . "Serial femtosecond crystallography of soluble proteins in lipidic cubic phase". United States. doi:10.1107/S2052252515013160. https://www.osti.gov/servlets/purl/1224055.
@article{osti_1224055,
title = {Serial femtosecond crystallography of soluble proteins in lipidic cubic phase},
author = {Fromme, Raimund and Ishchenko, Andrii and Metz, Markus and Chowdhury, Shatabdi Roy and Basu, Shibom and Boutet, Sébastien and Fromme, Petra and White, Thomas A. and Barty, Anton and Spence, John C. H. and Weierstall, Uwe and Liu, Wei and Cherezov, Vadim},
abstractNote = {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 shown 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.},
doi = {10.1107/S2052252515013160},
journal = {IUCrJ},
number = 5,
volume = 2,
place = {United States},
year = {Tue Aug 04 00:00:00 EDT 2015},
month = {Tue Aug 04 00:00:00 EDT 2015}
}

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Works referenced in this record:

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Gas dynamic virtual nozzle for generation of microscopic droplet streams
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