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Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

Journal article
Authors 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
Robert Dods
Rajiv Harimoorthy
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
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
G. Schertler
J. Standfuss
Published in Nature Communications
Volume 7
ISSN 2041-1723
Publication year 2016
Published at Department of Chemistry and Molecular Biology
Language en
Links dx.doi.org/10.1038/ncomms12314
Keywords x-ray-diffraction, protein-coupled receptors, coherent-light source, femtosecond crystallography, membrane-proteins, radiation-damage, macromolecular crystallography, structural alterations, photocycle, kinetics, bacteriorhodopsin, Science & Technology - Other Topics
Subject categories Chemical Sciences, Biological Sciences

Abstract

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.

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