Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
Description
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.
Date Created
The date the item was original created (prior to any relationship with the ASU Digital Repositories.)
2016-08-22
Agent
- Author (aut): Nogly, Przemyslaw
- Author (aut): Panneels, Valerie
- Author (aut): Nelson, Garrett
- Author (aut): Gati, Cornelius
- Author (aut): Kimura, Tetsunari
- Author (aut): Milne, Christopher
- Author (aut): Milathianaki, Despina
- Author (aut): Kubo, Minoru
- Author (aut): Wu, Wenting
- Author (aut): Conrad, Chelsie
- Author (aut): Coe, Jesse
- Author (aut): Bean, Richard
- Author (aut): Zhao, Yun
- Author (aut): Bath, Petra
- Author (aut): Dods, Robert
- Author (aut): Harimoorthy, Rajiv
- Author (aut): Beyerlein, Kenneth R.
- Author (aut): Rheinberger, Jan
- Author (aut): James, Daniel
- Author (aut): Deponte, Daniel
- Author (aut): Li, Chufeng
- Author (aut): Sala, Leonardo
- Author (aut): Williams, Garth J.
- Author (aut): Hunter, Mark S.
- Author (aut): Koglin, Jason E.
- Author (aut): Berntsen, Peter
- Author (aut): Nango, Eriko
- Author (aut): Iwata, So
- Author (aut): Chapman, Henry N.
- Author (aut): Fromme, Petra
- Author (aut): Frank, Matthias
- Author (aut): Abela, Rafael
- Author (aut): Boutet, Sebastien
- Author (aut): Barty, Anton
- Author (aut): White, Thomas A.
- Author (aut): Weierstall, Uwe
- Author (aut): Spence, John
- Author (aut): Neutze, Richard
- Author (aut): Schertler, Gebhard
- Author (aut): Standfuss, Jorg
- Contributor (ctb): College of Liberal Arts and Sciences
- Contributor (ctb): Department of Physics
- Contributor (ctb): Department of Chemistry and Biochemistry
- Contributor (ctb): Biodesign Institute
- Contributor (ctb): Applied Structural Discovery
- Contributor (ctb): School of Molecular Sciences