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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 n...

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Autores principales: Wu, Wenting, Nogly, Przemyslaw, Rheinberger, Jan, Kick, Leonhard M., Gati, Cornelius, Nelson, Garrett, Deupi, Xavier, Standfuss, Jörg, Schertler, Gebhard, Panneels, Valérie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498706/
https://www.ncbi.nlm.nih.gov/pubmed/26144230
http://dx.doi.org/10.1107/S2053230X15009966
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author Wu, Wenting
Nogly, Przemyslaw
Rheinberger, Jan
Kick, Leonhard M.
Gati, Cornelius
Nelson, Garrett
Deupi, Xavier
Standfuss, Jörg
Schertler, Gebhard
Panneels, Valérie
author_facet Wu, Wenting
Nogly, Przemyslaw
Rheinberger, Jan
Kick, Leonhard M.
Gati, Cornelius
Nelson, Garrett
Deupi, Xavier
Standfuss, Jörg
Schertler, Gebhard
Panneels, Valérie
author_sort Wu, Wenting
collection PubMed
description 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, 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.
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spelling pubmed-44987062015-07-28 Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser Wu, Wenting Nogly, Przemyslaw Rheinberger, Jan Kick, Leonhard M. Gati, Cornelius Nelson, Garrett Deupi, Xavier Standfuss, Jörg Schertler, Gebhard Panneels, Valérie Acta Crystallogr F Struct Biol Commun Iccbm15 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, 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. International Union of Crystallography 2015-06-27 /pmc/articles/PMC4498706/ /pubmed/26144230 http://dx.doi.org/10.1107/S2053230X15009966 Text en © Wu et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Iccbm15
Wu, Wenting
Nogly, Przemyslaw
Rheinberger, Jan
Kick, Leonhard M.
Gati, Cornelius
Nelson, Garrett
Deupi, Xavier
Standfuss, Jörg
Schertler, Gebhard
Panneels, Valérie
Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title_full Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title_fullStr Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title_full_unstemmed Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title_short Batch crystallization of rhodopsin for structural dynamics using an X-ray free-electron laser
title_sort batch crystallization of rhodopsin for structural dynamics using an x-ray free-electron laser
topic Iccbm15
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4498706/
https://www.ncbi.nlm.nih.gov/pubmed/26144230
http://dx.doi.org/10.1107/S2053230X15009966
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