Journal Article

PUBDB-2015-04296

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A novel inert crystal delivery medium for serial femtosecond crystallography

Conrad, C. E. ; Basu, S. ; James, D. ; Wang, D. ; Schaffer, A. ; Roy-Chowdhury, S. ; Zatsepin, N. A. ; Aquila, A.>Extern* ; Coe, J.>Extern* ; Gati, C.CFEL* ; Hunter, M. S. ; Koglin, J. E. ; Kupitz, C. ; Nelson, G. ; Subramanian, G. ; White, T.CFEL*DESY* ; Zhao, Y. ; Zook, J. ; Boutet, S.>Extern* ; Cherezov, V. ; Spence, J. C. H. ; Fromme, R.>Extern* ; Weierstall, U.>Extern* ; Fromme, P. (Corresponding author)>Extern*

2015
International Union of Crystallography (IUCr) Chester

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Please use a persistent id in citations: doi: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, femtosecondX-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 crystalladen agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 mg 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.

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Contributing Institute(s):

1. CFEL-Coherent X-Ray Imaging (FS-CFEL-1)

Research Program(s):

1. 6215 - Soft Matter, Health and Life Sciences (POF3-621) (POF3-621)

Experiment(s):

1. Experiments at CFEL

Appears in the scientific report 2015

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Database coverage:
; ; ; ; Current Contents - Physical, Chemical and Earth Sciences ; NCBI Molecular Biology Database ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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