2024-03-29T09:33:50Zhttps://keep.lib.asu.edu/oai/requestoai:keep.lib.asu.edu:node-1303182021-08-16T19:23:30Zoai_pmh:all130318
https://hdl.handle.net/2286/R.I.44351
Nogly, P., Panneels, V., Nelson, G., Gati, C., Kimura, T., Milne, C., . . . Standfuss, J. (2016). Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography. Nature Communications, 7, 12314. doi:10.1038/ncomms12314
10.1038/ncomms12314
2041-1723
http://rightsstatements.org/vocab/InC/1.0/
http://creativecommons.org/licenses/by/4.0
2016-08-22
9 pages
eng
Nogly, Przemyslaw
Panneels, Valerie
Nelson, Garrett
Gati, Cornelius
Kimura, Tetsunari
Milne, Christopher
Milathianaki, Despina
Kubo, Minoru
Wu, Wenting
Conrad, Chelsie
Coe, Jesse
Bean, Richard
Zhao, Yun
Bath, Petra
Dods, Robert
Harimoorthy, Rajiv
Beyerlein, Kenneth R.
Rheinberger, Jan
James, Daniel
Deponte, Daniel
Li, Chufeng
Sala, Leonardo
Williams, Garth J.
Hunter, Mark S.
Koglin, Jason E.
Berntsen, Peter
Nango, Eriko
Iwata, So
Chapman, Henry N.
Fromme, Petra
Frank, Matthias
Abela, Rafael
Boutet, Sebastien
Barty, Anton
White, Thomas A.
Weierstall, Uwe
Spence, John
Neutze, Richard
Schertler, Gebhard
Standfuss, Jorg
College of Liberal Arts and Sciences
Department of Physics
Department of Chemistry and Biochemistry
Biodesign Institute
Applied Structural Discovery
School of Molecular Sciences
The final version of this article, as published in Nature Communications, can be viewed online at: https://www.nature.com/articles/ncomms12314
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 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms 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 1 mg 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.
Text
Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography