A novel inert crystal delivery medium for serial femtosecond crystallography
Category
Published on
Type
journal-article
Author
Chelsie E. Conrad and Shibom Basu and Daniel James and Dingjie Wang and Alexander Schaffer and Shatabdi Roy-Chowdhury and Nadia A. Zatsepin and Andrew Aquila and Jesse Coe and Cornelius Gati and Mark S. Hunter and Jason E. Koglin and Christopher Kupitz and Garrett Nelson and Ganesh Subramanian and Thomas A. White and Yun Zhao and James Zook and Sébastien Boutet and Vadim Cherezov and John C. H. Spence and Raimund Fromme and Uwe Weierstall and Petra Fromme
Citation
Conrad, C.E. et al., 2015. A novel inert crystal delivery medium for serial femtosecond crystallography. IUCrJ, 2(4), pp.421–430. Available at: http://dx.doi.org/10.1107/s2052252515009811.
Abstract
Serial femtosecond crystallography (SFX) has opened a new era in crystallography by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 µg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.
DOI
Funding
NSF-STC Biology with X-ray Lasers (NSF-1231306)
