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Title: Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser

Rhodopsin is a membrane protein from the G protein-coupled receptor family. Together with its ligand retinal, it forms the visual pigment responsible for night vision. In order to perform ultrafast dynamics studies, a time-resolved serial femtosecond crystallography method is required owing to the nonreversible activation of rhodopsin. In such an approach, microcrystals in suspension are delivered into the X-ray pulses of an X-ray free-electron laser (XFEL) after a precise photoactivation delay. Here in this study, a millilitre batch production of high-density microcrystals was developed by four methodical conversion steps starting from known vapour-diffusion crystallization protocols: (i) screening the low-salt crystallization conditions preferred for serial crystallography by vapour diffusion, (ii) optimization of batch crystallization, (iii) testing the crystal size and quality using second-harmonic generation (SHG) imaging and X-ray powder diffraction and (iv) production of millilitres of rhodopsin crystal suspension in batches for serial crystallography tests; these crystals diffracted at an XFEL at the Linac Coherent Light Source using a liquid-jet setup.
Authors:
 [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1] ;  [1]
  1. Paul Scherrer Inst., Villigen (Switzerland). Lab. for Biomolecular Research
Publication Date:
Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section F, Structural Biology Communications
Additional Journal Information:
Journal Volume: 71; Journal Issue: 7; Journal ID: ISSN 2053-230X
Publisher:
International Union of Crystallography
Research Org:
Paul Scherrer Inst., Villigen (Switzerland). Lab. for Biomolecular Research
Sponsoring Org:
USDOE; National Institutes of Health (NIH); National Science Foundation (NSF)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; batch crystallization; GPCR; serial crystallography; FEL; dynamics
OSTI Identifier:
1345038

Wu, Wenting, Nogly, Przemyslaw, Rheinberger, Jan, Kick, Leonhard M., Gati, Cornelius, Nelson, Garrett, Deupi, Xavier, Standfuss, Jorg, Schertler, Gebhard, and Panneels, Valerie. Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser. United States: N. p., Web. doi:10.1107/s2053230x15009966.
Wu, Wenting, Nogly, Przemyslaw, Rheinberger, Jan, Kick, Leonhard M., Gati, Cornelius, Nelson, Garrett, Deupi, Xavier, Standfuss, Jorg, Schertler, Gebhard, & Panneels, Valerie. Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser. United States. doi:10.1107/s2053230x15009966.
Wu, Wenting, Nogly, Przemyslaw, Rheinberger, Jan, Kick, Leonhard M., Gati, Cornelius, Nelson, Garrett, Deupi, Xavier, Standfuss, Jorg, Schertler, Gebhard, and Panneels, Valerie. 2015. "Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser". United States. doi:10.1107/s2053230x15009966. https://www.osti.gov/servlets/purl/1345038.
@article{osti_1345038,
title = {Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser},
author = {Wu, Wenting and Nogly, Przemyslaw and Rheinberger, Jan and Kick, Leonhard M. and Gati, Cornelius and Nelson, Garrett and Deupi, Xavier and Standfuss, Jorg and Schertler, Gebhard and Panneels, Valerie},
abstractNote = {Rhodopsin is a membrane protein from the G protein-coupled receptor family. Together with its ligand retinal, it forms the visual pigment responsible for night vision. In order to perform ultrafast dynamics studies, a time-resolved serial femtosecond crystallography method is required owing to the nonreversible activation of rhodopsin. In such an approach, microcrystals in suspension are delivered into the X-ray pulses of an X-ray free-electron laser (XFEL) after a precise photoactivation delay. Here in this study, a millilitre batch production of high-density microcrystals was developed by four methodical conversion steps starting from known vapour-diffusion crystallization protocols: (i) screening the low-salt crystallization conditions preferred for serial crystallography by vapour diffusion, (ii) optimization of batch crystallization, (iii) testing the crystal size and quality using second-harmonic generation (SHG) imaging and X-ray powder diffraction and (iv) production of millilitres of rhodopsin crystal suspension in batches for serial crystallography tests; these crystals diffracted at an XFEL at the Linac Coherent Light Source using a liquid-jet setup.},
doi = {10.1107/s2053230x15009966},
journal = {Acta Crystallographica. Section F, Structural Biology Communications},
number = 7,
volume = 71,
place = {United States},
year = {2015},
month = {6}
}