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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Bibliographic Details
Published in:IUCrJ 2015-09, Vol.2 (Pt 5), p.545-551
Main Authors: 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
Format: Article
Language:English
Subjects:
Crystallography
Data acquisition
Femtosecond
Free electron lasers
Injectors
lipidic cubic phase
Liquid crystal polymers
Methods
Microcrystals
PARTICLE ACCELERATORS
Proteins
Research Papers
serial femtosecond crystallography
Serials
soluble protein
Structure
X-ray free-electron laser
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Summary: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.
ISSN:2052-2525
2052-2525
DOI:10.1107/s2052252515013160