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.
protein crystallography
MACROMOLECULAR
Chemistry
CRYSTALLIZATION
Multidisciplinary
DAMAGE
lipidic cubic phases
FREE-ELECTRON
CRYSTALLOGRAPHY
XFEL
FEMTOSECOND CRYSTALLOGRAPHY
PROTEIN-STRUCTURE DETERMINATION
LASER
Materials Science
Multidisciplinary
Crystallography
MEMBRANE-PROTEINS
bacteriorhodopsin
RESOLUTION
X-RAY CRYSTALLOGRAPHY
SOFTWARE
Author
P. Nogly
D. James
D. J. Wang
T. A. White
N. Zatsepin
A. Shilova
G. Nelson
H. G. Liu
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
Chemical Sciences
DOI
10.1107/s2052252514026487
PubMed
25866654