Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
Journal article, 2016
Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 angstrom resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.
femtosecond crystallography
coherent-light source
macromolecular crystallography
membrane-proteins
Science & Technology - Other Topics
kinetics
bacteriorhodopsin
x-ray-diffraction
radiation-damage
structural alterations
photocycle
protein-coupled receptors
Author
P. Nogly
V. Panneels
G. Nelson
C. Gati
T. Kimura
C. Milne
D. Milathianaki
M. Kubo
W. T. Wu
C. Conrad
J. Coe
R. Bean
Y. Zhao
Petra Båth
University of Gothenburg
Robert Dods
University of Gothenburg
Rajiv Harimoorthy
University of Gothenburg
K. R. Beyerlein
J. Rheinberger
D. James
D. DePonte
C. F. Li
L. Sala
G. J. Williams
M. S. Hunter
J. E. Koglin
Peter Berntsen
University of Gothenburg
E. Nango
S. Iwata
H. N. Chapman
P. Fromme
M. Frank
R. Abela
S. Boutet
A. Barty
T. A. White
U. Weierstall
J. Spence
Richard Neutze
University of Gothenburg
G. Schertler
J. Standfuss
Nature Communications
2041-1723 (ISSN) 20411723 (eISSN)
Vol. 7Subject Categories
Biological Sciences
Chemical Sciences
DOI
10.1038/ncomms12314