A novel inert crystal delivery medium for serial femtosecond crystallography

Chelsie E. Conrad, Shibom Basu, Daniel James, Dingjie Wang, Alexander Schaffer, Shatabdi Roy-Chowdhury, Nadia 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 CherezovJohn Spence, Raimund Fromme, Uwe Weierstall, Petra Fromme

Research output: Contribution to journalArticle

74 Scopus citations

Abstract

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°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.

Original languageEnglish (US)
Pages (from-to)421-430
Number of pages10
JournalIUCrJ
Volume2
DOIs
StatePublished - Jul 1 2015

Keywords

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

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry
  • Materials Science(all)
  • Condensed Matter Physics

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  • Cite this

    Conrad, C. E., Basu, S., James, D., Wang, D., Schaffer, A., Roy-Chowdhury, S., Zatsepin, N., Aquila, A., Coe, J., Gati, C., Hunter, M. S., Koglin, J. E., Kupitz, C., Nelson, G., Subramanian, G., White, T. A., Zhao, Y., Zook, J., Boutet, S., ... Fromme, P. (2015). A novel inert crystal delivery medium for serial femtosecond crystallography. IUCrJ, 2, 421-430. https://doi.org/10.1107/S2052252515009811