A novel inert crystal delivery medium for serial femtosecond crystallography

作者：Conrad Chelsie E; Basu Shibom; James Daniel; Wang Dingjie; Schaffer Alexander; Roy Chowdhury Shatabdi; Zatsepin Nadia A; Aquila Andrew; Coe Jesse; Gati Cornelius; Hunter Mark S; Koglin Jason E; Kupitz Christopher; Nelson Garrett; Subramanian Ganesh; White Thomas A; Zhao Yun; Zook James; Boutet Sebastien; Cherezov Vadim; Spence John C H; Fromme Raimund; Weierstall Uwe; Fromme Petra^*

来源：IUCrJ, 2015, 2(4): 421-430.

DOI：10.1107/S2052252515009811

• serial femtosecond crystallography
• viscous crystal delivery
• protein complexes
• membrane proteins
• femtosecond studies
• nanocrystals
• coherent X-ray diffractive imaging
• free-electron laser

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 degrees C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 angstrom resolution using 300 mu g 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.

• 出版日期2015-7