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Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

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 b...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4547822/
https://www.ncbi.nlm.nih.gov/pubmed/26306196
http://dx.doi.org/10.1107/S2052252515013160
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author 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
author_facet 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
author_sort Fromme, Raimund
collection PubMed
description 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.
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spelling pubmed-45478222015-08-24 Serial femtosecond crystallography of soluble proteins in lipidic cubic phase 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 IUCrJ Research Papers 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. International Union of Crystallography 2015-08-04 /pmc/articles/PMC4547822/ /pubmed/26306196 http://dx.doi.org/10.1107/S2052252515013160 Text en © Raimund Fromme et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Papers
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
title Serial femtosecond crystallography of soluble proteins in lipidic cubic phase
title_full Serial femtosecond crystallography of soluble proteins in lipidic cubic phase
title_fullStr Serial femtosecond crystallography of soluble proteins in lipidic cubic phase
title_full_unstemmed Serial femtosecond crystallography of soluble proteins in lipidic cubic phase
title_short Serial femtosecond crystallography of soluble proteins in lipidic cubic phase
title_sort serial femtosecond crystallography of soluble proteins in lipidic cubic phase
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4547822/
https://www.ncbi.nlm.nih.gov/pubmed/26306196
http://dx.doi.org/10.1107/S2052252515013160
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