Lipidic cubic phase serial millisecond crystallography using synchrotron radiation

Przemyslaw Nogly (1) , Daniel James (2) , Dingjie Wang (2) , Thomas A. White (3) , Nadia Zatsepin (2) , Anastasya Shilova (4) , Garrett Nelson (2) , Haiguang Liu (2) , Linda Johansson (5) , Michael Heymann (3) , Kathrin Jaeger (1) , Markus Metz (6, 3) , Cecilia Wickstrand (7) , Wenting Wu (1) , Petra Bath (7) , Peter Berntsen (7) , Dominik Oberthuer (6, 3) , Valerie Panneels (1) , Vadim Cherezov (5) , Henry Chapman (3, 8) , Gebhard Schertler (9, 1) , Richard Neutze (7) , John Spence (10) , Isabel Moraes (11, 12, 13) , Manfred Burghammer (14, 4) , Joerg Standfuss (1) , Uwe Weierstall (2)

1 Paul Scherrer Inst, Lab Biomol Res, CH-5232 Villigen, Switzerland
2 ASU - Arizona State University [Tempe]
3 DESY, Ctr Free Electron Laser Sci, D-22607 Hamburg, Germany
4 ESRF - European Synchrotron Radiation Facility
5 The Scripps Research Institute [La Jolla, San Diego]
6 Ctr Ultrafast Imaging, D-22607 Hamburg, Germany
7 GU - Göteborgs Universitet = University of Gothenburg
8 Univ Hamburg, Dept Phys, D-22607 Hamburg, Germany
9 D-BIOL - Department of Biology [ETH Zürich]
10 Department of Physics [Tempe]
11 DIAMOND Light source
12 Rutherford Appleton Lab, Didcot OX11 0FA, Oxon, England
13 Univ London Imperial Coll Sci Technol & Med, Dept Life Sci, London, England
14 UGENT - Universiteit Gent = Ghent University

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 angstrom. 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

Mots clés: XFEL; bacteriorhodopsin; lipidic cubic phases; protein crystallography

Domaines: Chimie

Przemyslaw Nogly, Daniel James, Dingjie Wang, Thomas A. White, Nadia Zatsepin, et al.. Lipidic cubic phase serial millisecond crystallography using synchrotron radiation. International Union of Crystallography journal, 2015, 2, pp.168-176. ⟨10.1107/S2052252514026487⟩. ⟨hal-01575306⟩