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

• DOI: 10.1016/j.str.2017.07.002
• Publication Date: 2017-08-03T20:16:44Z
• Authors: Robert Dods; Petra Båth; David Arnlund; Kenneth R. Beyerlein; Garrett Nelson; Mengling Liang; Rajiv Harimoorthy; Peter Berntsen; Erik Malmerberg; Linda Johansson; Rebecka Andersson; Robert Bosman; Sergio Carbajo; Elin Claesson; Chelsie E. Conrad; Peter Dahl; Greger Hammarin; Mark S. Hunter; Chufeng Li; Stella Lisova; Despina Milathianaki; Joseph Robinson; Cecilia Safari; Amit Sharma; Garth Williams; Cecilia Wickstrand; Oleksandr Yefanov; Jan Davidsson; Daniel P. DePonte; Anton Barty; Gisela Brändén; Richard Neutze
• Keywords: info:eu-repo/classification/ddc/570; Models, Molecular; 0301 basic medicine; Hyphomicrobiaceae; 03 medical and health sciences; Bacterial Proteins; Protein Conformation; Membrane Proteins; Photosynthesis; Crystallography, X-Ray

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.