A novel inert crystal delivery medium for serial femtosecond crystallography

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, Sébastien; Cherezov, Vadim; Spence, John C. H.; Fromme, Raimund; Weierstall, Uwe; Fromme, Petra

doi:10.1107/S2052252515009811

A novel inert crystal delivery medium for serial femtosecond crystallography

Journal Article · Tue Jun 30 00:00:00 EDT 2015 · IUCrJ

DOI:https://doi.org/10.1107/S2052252515009811· OSTI ID:1208832

Conrad, Chelsie E. ^[1]; Basu, Shibom ^[1]; James, Daniel ^[1]; Wang, Dingjie ^[2]; Schaffer, Alexander ^[1]; Roy-Chowdhury, Shatabdi ^[1]; Zatsepin, Nadia A. ^[1]; Aquila, Andrew ^[3]; Coe, Jesse ^[1]; Gati, Cornelius ^[4]; Hunter, Mark S. ^[3]; Koglin, Jason E. ^[5]; Kupitz, Christopher ^[6]; Nelson, Garrett ^[1]; Subramanian, Ganesh ^[1]; White, Thomas A. ^[4]; Zhao, Yun ^[4]; Zook, James ^[1]; Boutet, Sébastien ^[5]; Cherezov, Vadim ^[7] more »

Arizona State Univ., Tempe, AZ (United States); The Biodesign Inst., Tempe, AZ (United States)
Arizona State Univ., Tempe, AZ (United States)
SLAC National Accelerator Laboratory, Menlo Park, CA (United States)
Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Univ. of Wisconsin-Milwaukee, Milwaukee, WI (United States); The Biodesign Inst., Tempe, AZ (United States)
Univ. of Southern California, Los Angeles, CA (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)

Grant/Contract Number:: AC03-76SF00515; SC0010575

OSTI ID:: 1208832

Alternate ID(s):: OSTI ID: 1261172

Journal Information:: IUCrJ, Journal Name: IUCrJ Journal Issue: 4 Vol. 2; ISSN 2052-2525; ISSN IUCRAJ

Publisher:: International Union of CrystallographyCopyright Statement

Country of Publication:: United States

Language:: English

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

A novel inert crystal delivery medium for serial femtosecond crystallography

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