Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
Artikel i vetenskaplig tidskrift, 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

Författare

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

Göteborgs universitet

Robert Dods

Göteborgs universitet

Rajiv Harimoorthy

Göteborgs universitet

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

Göteborgs universitet

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

Göteborgs universitet

G. Schertler

J. Standfuss

Nature Communications

2041-1723 (ISSN)

Vol. 7

Ämneskategorier

Biologiska vetenskaper

Kemi

DOI

10.1038/ncomms12314