Lipidic cubic phase serial millisecond crystallography using synchrotron radiation

Przemyslaw Nogly; Daniel James; Dingjie Wang; Thomas A White; Nadia Zatsepin; Anastasya Shilova; Garrett Nelson; Haiguang Liu; Linda Johansson; Michael Heymann; Kathrin Jaeger; Markus Metz; Cecilia Wickstrand; Wenting Wu; Petra Båth; Peter Berntsen; Dominik Oberthuer; Valerie Panneels; Vadim Cherezov; Henry Chapman; Gebhard Schertler; Richard Neutze; John Spence; Isabel Moraes; Manfred Burghammer; Joerg Standfuss; Uwe Weierstall

doi:10.1107/S2052252514026487

Lipidic cubic phase serial millisecond crystallography using synchrotron radiation

IUCrJ. 2015 Jan 27;2(Pt 2):168-76. doi: 10.1107/S2052252514026487. eCollection 2015 Mar 1.

Authors

Przemyslaw Nogly¹, Daniel James², Dingjie Wang², Thomas A White³, Nadia Zatsepin², Anastasya Shilova⁴, Garrett Nelson², Haiguang Liu², Linda Johansson⁵, Michael Heymann³, Kathrin Jaeger¹, Markus Metz⁶, Cecilia Wickstrand⁷, Wenting Wu¹, Petra Båth⁷, Peter Berntsen⁷, Dominik Oberthuer⁶, Valerie Panneels¹, Vadim Cherezov⁵, Henry Chapman⁸, Gebhard Schertler⁹, Richard Neutze⁷, John Spence², Isabel Moraes¹⁰, Manfred Burghammer¹¹, Joerg Standfuss¹, Uwe Weierstall²

Affiliations

¹ Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
² Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
³ Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Hamburg 22607, Germany.
⁴ European Synchrotron Radiation Facility, Grenoble Cedex 9, F-38043, France.
⁵ Department of Integrative Structural and Computational Biology, Scripps Research Institute, La Jolla, California USA.
⁶ Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Hamburg 22607, Germany ; Centre for Ultrafast Imaging, Hamburg 22607, Germany.
⁷ Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden.
⁸ Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Hamburg 22607, Germany ; Department of Physics, University of Hamburg, Hamburg 22607, Germany.
⁹ Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland ; Deparment of Biology, ETH Zurich, Zürich 8093, Switzerland.
¹⁰ Membrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation Campus, Chilton, Oxfordshire OX11 0DE, England ; Department of Life Sciences, Imperial College London, London, England ; Research Complex at Harwell Rutherford, Appleton Laboratory, Harwell, Didcot, Oxfordshire OX11 0FA, England.
¹¹ European Synchrotron Radiation Facility, Grenoble Cedex 9, F-38043, France ; Department of Analytical Chemistry, Ghent University, Ghent B-9000, Belgium.

Abstract

Lipidic cubic phases (LCPs) have emerged as successful matrixes for the crystallization of membrane proteins. Moreover, the viscous LCP also provides a highly effective delivery medium for serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs). Here, the adaptation of this technology to perform serial millisecond crystallography (SMX) at more widely available synchrotron microfocus beamlines is described. Compared with conventional microcrystallography, LCP-SMX eliminates the need for difficult handling of individual crystals and allows for data collection at room temperature. The technology is demonstrated by solving a structure of the light-driven proton-pump bacteriorhodopsin (bR) at a resolution of 2.4 Å. The room-temperature structure of bR is very similar to previous cryogenic structures but shows small yet distinct differences in the retinal ligand and proton-transfer pathway.

Keywords: XFEL; bacteriorhodopsin; lipidic cubic phases; protein crystallography.

Abstract

Grants and funding