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Lipidic cubic phase serial millisecond crystallography using synchrotron radiation

DOI: 10.1107/S2052252514026487 DOI Help
PMID: 25866654 PMID Help

Authors: Przemyslaw Nogly (Instituto de Tecnologia Química e Biológica) , Daniel James (Arizona State University) , Dingjie Wang (Arizona State University) , Thomas A. White (DESY) , Nadia Zatsepin (Arizona State University) , Anastasya Shilova (Arizona State University) , Garrett Nelson (Arizona State University) , Haiguang Liu (Arizona State University) , Linda Johansson (Department of Integrative Structural and Computational Biology, Scripps Research Institute) , Michael Heymann (Deutsches Elektronen-Synchrotron DESY) , Kathrin Jaeger (Laboratory for Biomolecular Research, Switzerland) , Markus Metz (Deutsches Elektronen-Synchrotron DESY) , Cecilia Wickstrand (University of Gothenburg) , Wenting Wu (Laboratory for Biomolecular Research, Switzerland) , Petra Båth (Department of Chemistry and Molecular Biology, University of Gothenburg) , Peter Berntsen (Department of Chemistry and Molecular Biology, University of Gothenburg) , Dominik Oberthuer (Deutsches Elektronen-Synchrotron DESY) , Valerie Panneels (Laboratory for Biomolecular Research, Switzerland) , Vadim Cherezov (Department of Integrative Structural and Computational Biology, Scripps Research Institute) , Isabel Moraes (Diamond Light Source; Imperial College London; Imperial College London) , Henry Chapman (Deutsches Elektronen-Synchrotron DESY; University of Hamburg) , Gebhard Schertler (Paul Scherrer Institute; ETH Zurich) , Richard Neutze (University of Gothenburg) , John Spence (Arizona State University) , Manfred Burghammer (European Synchrotron Radiation Facility; Ghent University) , Joerg Standfuss (Paul Scherrer Institute) , Uwe Weierstall (Arizona State University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Iucrj , VOL 2 , PAGES 168 - 176

State: Published (Approved)
Published: March 2015

Open Access Open Access

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.

Journal Keywords: Lipidic Cubic Phases; Protein Crystallography; Bacteriorhodopsin; Xfel.

Subject Areas: Biology and Bio-materials, Medicine

Diamond Offline Facilities: Membrane Protein Laboratory (MPL)

Other Facilities: ESRF ID02 beamline and ID13 PXI beamline Swiss Light Source