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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
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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. |
format | Online Article Text |
id | pubmed-4498706 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
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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