A novel inert crystal delivery medium for serial femtosecond crystallography
| Autor: | Chelsie E. Conrad, Shibom Basu, Daniel James, Dingjie Wang, Alexander Schaffer, Shatabdi Roy-Chowdhury, Nadia A. Zatsepin, Andrew Aquila, Jesse Coe, Cornelius Gati, Mark S. Hunter, Jason E. Koglin, Christopher Kupitz, Garrett Nelson, Ganesh Subramanian, Thomas A. White, Yun Zhao, James Zook, Sébastien Boutet, Vadim Cherezov, John C. H. Spence, Raimund Fromme, Uwe Weierstall, Petra Fromme |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: | |
| Zdroj: | IUCrJ, Vol 2, Iss 4, Pp 421-430 (2015) |
| Druh dokumentu: | article |
| ISSN: | 2052-2525 20522525 |
| DOI: | 10.1107/S2052252515009811 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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