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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...

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Autores principales: Rodrigues, Matthew J., Casadei, Cecilia M., Weinert, Tobias, Panneels, Valerie, Schertler, Gebhard F. X.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2023
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.
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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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