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

Journal article
Authors Robert Dods
Petra Båth
David Arnlund
Kenneth R Beyerlein
Garrett Nelson
Mengling Liang
Rajiv Harimoorthy
Peter Berntsen
Erik Malmerberg
Linda C 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
Published in Structure
Volume 25
Issue 9
Pages 1461-1468
ISSN 1878-4186
Publication year 2017
Published at Department of Chemistry and Molecular Biology
Pages 1461-1468
Language en
Links dx.doi.org/10.1016/j.str.2017.07.00...
www.ncbi.nlm.nih.gov/entrez/query.f...
Subject categories Structural Biology, Biochemistry, Molecular biophysics

Abstract

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.

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