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Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect
Rhodopsin is a G-protein-coupled receptor that detects light and initiates the intracellular signalling cascades that underpin vertebrate vision. Light sensitivity is achieved by covalent linkage to 11-cis retinal, which isomerizes upon photo-absorption. Serial femtosecond crystallography data colle...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9986800/ https://www.ncbi.nlm.nih.gov/pubmed/36876432 http://dx.doi.org/10.1107/S2059798323000931 |
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author | Rodrigues, Matthew J. Casadei, Cecilia M. Weinert, Tobias Panneels, Valerie Schertler, Gebhard F. X. |
author_facet | Rodrigues, Matthew J. Casadei, Cecilia M. Weinert, Tobias Panneels, Valerie Schertler, Gebhard F. X. |
author_sort | Rodrigues, Matthew J. |
collection | PubMed |
description | Rhodopsin is a G-protein-coupled receptor that detects light and initiates the intracellular signalling cascades that underpin vertebrate vision. Light sensitivity is achieved by covalent linkage to 11-cis retinal, which isomerizes upon photo-absorption. Serial femtosecond crystallography data collected from rhodopsin microcrystals grown in the lipidic cubic phase were used to solve the room-temperature structure of the receptor. Although the diffraction data showed high completeness and good consistency to 1.8 Å resolution, prominent electron-density features remained unaccounted for throughout the unit cell after model building and refinement. A deeper analysis of the diffraction intensities uncovered the presence of a lattice-translocation defect (LTD) within the crystals. The procedure followed to correct the diffraction intensities for this pathology enabled the building of an improved resting-state model. The correction was essential to both confidently model the structure of the unilluminated state and interpret the light-activated data collected after photo-excitation of the crystals. It is expected that similar cases of LTD will be observed in other serial crystallography experiments and that correction will be required in a variety of systems. |
format | Online Article Text |
id | pubmed-9986800 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-99868002023-03-07 Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect Rodrigues, Matthew J. Casadei, Cecilia M. Weinert, Tobias Panneels, Valerie Schertler, Gebhard F. X. Acta Crystallogr D Struct Biol Research Papers Rhodopsin is a G-protein-coupled receptor that detects light and initiates the intracellular signalling cascades that underpin vertebrate vision. Light sensitivity is achieved by covalent linkage to 11-cis retinal, which isomerizes upon photo-absorption. Serial femtosecond crystallography data collected from rhodopsin microcrystals grown in the lipidic cubic phase were used to solve the room-temperature structure of the receptor. Although the diffraction data showed high completeness and good consistency to 1.8 Å resolution, prominent electron-density features remained unaccounted for throughout the unit cell after model building and refinement. A deeper analysis of the diffraction intensities uncovered the presence of a lattice-translocation defect (LTD) within the crystals. The procedure followed to correct the diffraction intensities for this pathology enabled the building of an improved resting-state model. The correction was essential to both confidently model the structure of the unilluminated state and interpret the light-activated data collected after photo-excitation of the crystals. It is expected that similar cases of LTD will be observed in other serial crystallography experiments and that correction will be required in a variety of systems. International Union of Crystallography 2023-02-27 /pmc/articles/PMC9986800/ /pubmed/36876432 http://dx.doi.org/10.1107/S2059798323000931 Text en © Matthew J. Rodrigues et al. 2023 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Rodrigues, Matthew J. Casadei, Cecilia M. Weinert, Tobias Panneels, Valerie Schertler, Gebhard F. X. Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title | Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title_full | Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title_fullStr | Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title_full_unstemmed | Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title_short | Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
title_sort | correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9986800/ https://www.ncbi.nlm.nih.gov/pubmed/36876432 http://dx.doi.org/10.1107/S2059798323000931 |
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