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

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

doi:10.1016/j.str.2017.07.002

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.

Authors

Robert Dods¹, Petra Båth¹, David Arnlund¹, Kenneth R Beyerlein², Garrett Nelson³, Mengling Liang⁴, Rajiv Harimoorthy¹, Peter Berntsen⁵, Erik Malmerberg⁶, Linda Johansson⁷, Rebecka Andersson¹, Robert Bosman¹, Sergio Carbajo⁴, Elin Claesson¹, Chelsie E Conrad³, Peter Dahl¹, Greger Hammarin¹, Mark S Hunter⁴, Chufeng Li³, Stella Lisova³, Despina Milathianaki⁴, Joseph Robinson⁴, Cecilia Safari¹, Amit Sharma¹, Garth Williams⁴, Cecilia Wickstrand¹, Oleksandr Yefanov², Jan Davidsson⁸, Daniel P DePonte², Anton Barty², Gisela Brändén⁹, Richard Neutze¹⁰

Affiliations

¹ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden.
² Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany.
³ Department of Physics, Arizona State University, Tempe, AZ, USA.
⁴ Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, Menlo Park, CA, USA.
⁵ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, La Trobe Institute for Molecular Science, La Trobe University, Melbourne 3086, Australia.
⁶ Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA 94720, USA.
⁷ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden; The Bridge Institute, Department of Chemistry, University of Southern California, Los Angeles, CA 90089-3303, USA.
⁸ Department of Photochemistry and Molecular Science, Uppsala University, Uppsala, Sweden.
⁹ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden. Electronic address: gisela.branden@gu.se.
¹⁰ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden. Electronic address: richard.neutze@gu.se.

PMID: 28781082
DOI: 10.1016/j.str.2017.07.002

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 (RC_vir). We first crushed these crystals prior to multiple rounds of microseeding. Each cycle of microseeding facilitated improvements in the RC_vir 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

Substances

Bacterial Proteins
Membrane Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding