A novel inert crystal delivery medium for serial femtosecond crystallography Academic Article uri icon

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, 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.

authors

  • 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 CH
  • Fromme, Raimund
  • Weierstall, Uwe
  • Fromme, Petra

publication date

  • 2015

published in