Description
Experimental Technique/Method:X-RAY DIFFRACTION
Resolution:2.3
Classification:PROTON TRANSPORT
Release Date:2016-08-31
Deposition Date:2016-04-06
Revision Date:2018-01-24
Molecular Weight:28341.51
Macromolecule Type:Protein
Residue Count:227
Atom Site Count:1792
DOI:10.2210/pdb5j7a/pdb
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 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.
Resolution:2.3
Classification:PROTON TRANSPORT
Release Date:2016-08-31
Deposition Date:2016-04-06
Revision Date:2018-01-24
Molecular Weight:28341.51
Macromolecule Type:Protein
Residue Count:227
Atom Site Count:1792
DOI:10.2210/pdb5j7a/pdb
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 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 made available | 2016 |
|---|---|
| Publisher | RCSB-PDB |