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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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
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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. |
format | Online Article Text |
id | pubmed-4547822 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
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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