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Publication Abstract from PubMed

Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy 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 A resolution using 300 microg 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.

A novel inert crystal delivery medium for serial femtosecond crystallography.,Conrad CE, Basu S, James D, Wang D, Schaffer A, Roy-Chowdhury S, Zatsepin NA, Aquila A, Coe J, Gati C, Hunter MS, Koglin JE, Kupitz C, Nelson G, Subramanian G, White TA, Zhao Y, Zook J, Boutet S, Cherezov V, Spence JC, Fromme R, Weierstall U, Fromme P IUCrJ. 2015 Jun 30;2(Pt 4):421-30. doi: 10.1107/S2052252515009811. eCollection, 2015 Jul 1. PMID:26177184^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.