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)

Vol. 7

Subject Categories

Biological Sciences

Chemical Sciences

DOI

10.1038/ncomms12314

More information

Created

10/10/2017