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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:
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 = {2015},
month = {8}
}

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Cited by: 20 works
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    Works referencing / citing this record:

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    Enabling membrane protein structure and dynamics with X-ray free electron lasers
    journal, August 2014


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    journal, November 2012


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    journal, September 2008

    • DePonte, D. P.; Weierstall, U.; Schmidt, K.
    • Journal of Physics D: Applied Physics, Vol. 41, Issue 19, Article No. 195505
    • DOI: 10.1088/0022-3727/41/19/195505

    Serial Femtosecond Crystallography of G Protein-Coupled Receptors
    journal, December 2013


    Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography
    journal, February 2014

    • Weierstall, Uwe; James, Daniel; Wang, Chong
    • Nature Communications, Vol. 5, Issue 1
    • DOI: 10.1038/ncomms4309

    Preparation of microcrystals in lipidic cubic phase for serial femtosecond crystallography
    journal, August 2014


    Grease matrix as a versatile carrier of proteins for serial crystallography
    journal, November 2014

    • Sugahara, Michihiro; Mizohata, Eiichi; Nango, Eriko
    • Nature Methods, Vol. 12, Issue 1
    • DOI: 10.1038/nmeth.3172

    Microcrystallization techniques for serial femtosecond crystallography using photosystem II from Thermosynechococcus elongatus as a model system
    journal, July 2014

    • Kupitz, Christopher; Grotjohann, Ingo; Conrad, Chelsie E.
    • Philosophical Transactions of the Royal Society B: Biological Sciences, Vol. 369, Issue 1647
    • DOI: 10.1098/rstb.2013.0316

    Lipidic phase membrane protein serial femtosecond crystallography
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    • Johansson, Linda C.; Arnlund, David; White, Thomas A.
    • Nature Methods, Vol. 9, Issue 3
    • DOI: 10.1038/nmeth.1867

    Protein crystal structure obtained at 2.9 A resolution from injecting bacterial cells into an X-ray free-electron laser beam
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    • Sawaya, M. R.; Cascio, D.; Gingery, M.
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    • Chapman, Henry N.; Fromme, Petra; Barty, Anton
    • Nature, Vol. 470, Issue 7332, p. 73-77
    • DOI: 10.1038/nature09750

    High-Resolution Protein Structure Determination by Serial Femtosecond Crystallography
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    • Chapman, Henry N.; Barty, Anton; Bogan, Michael J.
    • Nature Physics, Vol. 2, Issue 12
    • DOI: 10.1038/nphys461