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Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

Authors :
Raimund Fromme
Andrii Ishchenko
Markus Metz
Shatabdi Roy Chowdhury
Shibom Basu
Sébastien Boutet
Petra Fromme
Thomas A. White
Anton Barty
John C. H. Spence
Uwe Weierstall
Wei Liu
Vadim Cherezov
Source :
IUCrJ, Vol 2, Iss 5, Pp 545-551 (2015)
Publication Year :
2015
Publisher :
International Union of Crystallography, 2015.

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.

Details

Language :
English
ISSN :
20522525
Volume :
2
Issue :
5
Database :
Directory of Open Access Journals
Journal :
IUCrJ
Publication Type :
Academic Journal
Accession number :
edsdoj.b6aa19b247f3a5c91329a740b0a7
Document Type :
article
Full Text :
https://doi.org/10.1107/S2052252515013160