From Macrocrystals to Microcrystals: A Strategy for Membrane Protein Serial Crystallography

Structure. 2017 Sep 5;25(9):1461-1468.e2. doi: 10.1016/j.str.2017.07.002. Epub 2017 Aug 3.


Serial protein crystallography was developed at X-ray free-electron lasers (XFELs) and is now also being applied at storage ring facilities. Robust strategies for the growth and optimization of microcrystals are needed to advance the field. Here we illustrate a generic strategy for recovering high-density homogeneous samples of microcrystals starting from conditions known to yield large (macro) crystals of the photosynthetic reaction center of Blastochloris viridis (RCvir). We first crushed these crystals prior to multiple rounds of microseeding. Each cycle of microseeding facilitated improvements in the RCvir serial femtosecond crystallography (SFX) structure from 3.3-Å to 2.4-Å resolution. This approach may allow known crystallization conditions for other proteins to be adapted to exploit novel scientific opportunities created by serial crystallography.

Keywords: X-ray free-electron laser; microcrystallography; photosynthetic reaction center; serial femtosecond crystallography.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Crystallography, X-Ray
  • Hyphomicrobiaceae / chemistry
  • Hyphomicrobiaceae / metabolism*
  • Membrane Proteins / chemistry*
  • Models, Molecular
  • Photosynthesis
  • Protein Conformation


  • Bacterial Proteins
  • Membrane Proteins