Lipidic cubic phase serial millisecond crystallography using synchrotron radiation
Journal article, 2015
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.
Crystallography
MEMBRANE-PROTEINS
lipidic cubic phases
protein crystallography
CRYSTALLIZATION
X-RAY CRYSTALLOGRAPHY
bacteriorhodopsin
CRYSTALLOGRAPHY
Multidisciplinary
PROTEIN-STRUCTURE DETERMINATION
Chemistry
DAMAGE
LASER
FREE-ELECTRON
FEMTOSECOND CRYSTALLOGRAPHY
XFEL
RESOLUTION
Multidisciplinary
Materials Science
MACROMOLECULAR
SOFTWARE
Author
P. Nogly
D. James
D. J. Wang
T. A. White
N. Zatsepin
A. Shilova
G. Nelson
H. G. Liu
Karl-Åke L Johansson
M. Heymann
K. Jaeger
M. Metz
C. Wickstrand
W. T. Wu
P. Bath
Peter Berntsen
University of Gothenburg
D. Oberthuer
V. Panneels
V. Cherezov
H. Chapman
G. Schertler
Richard Neutze
University of Gothenburg
J. Spence
I. Moraes
M. Burghammer
J. Standfuss
U. Weierstall
IUCrJ
2052-2525 (eISSN)
Vol. 2 168-176 Part: 2Subject Categories (SSIF 2011)
Chemical Sciences
DOI
10.1107/s2052252514026487
PubMed
25866654