Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin

Abstract Mutations in the G protein-coupled receptor (GPCR) rhodopsin are a common cause of autosomal dominant retinitis pigmentosa, a blinding disease. Rhodopsin self-associates in the membrane, and the purified monomeric apo-protein opsin dimerizes in vitro as it transitions from detergent micelle...

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Autores principales:	George Khelashvili, Anoop Narayana Pillai, Joon Lee, Kalpana Pandey, Alexander M. Payne, Zarek Siegel, Michel A. Cuendet, Tylor R. Lewis, Vadim Y. Arshavsky, Johannes Broichhagen, Joshua Levitz, Anant K. Menon
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dd6f556860b34a2ea32087d0f5c5847f
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spelling	oai:doaj.org-article:dd6f556860b34a2ea32087d0f5c5847f2021-12-02T15:52:25ZUnusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin10.1038/s41598-021-90039-32045-2322https://doaj.org/article/dd6f556860b34a2ea32087d0f5c5847f2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90039-3https://doaj.org/toc/2045-2322Abstract Mutations in the G protein-coupled receptor (GPCR) rhodopsin are a common cause of autosomal dominant retinitis pigmentosa, a blinding disease. Rhodopsin self-associates in the membrane, and the purified monomeric apo-protein opsin dimerizes in vitro as it transitions from detergent micelles to reconstitute into a lipid bilayer. We previously reported that the retinitis pigmentosa-linked F220C opsin mutant fails to dimerize in vitro, reconstituting as a monomer. Using fluorescence-based assays and molecular dynamics simulations we now report that whereas wild-type and F220C opsin display distinct dimerization propensities in vitro as previously shown, they both dimerize in the plasma membrane of HEK293 cells. Unexpectedly, molecular dynamics simulations show that F220C opsin forms an energetically favored dimer in the membrane when compared with the wild-type protein. The conformation of the F220C dimer is unique, with transmembrane helices 5 and 6 splayed apart, promoting widening of the intracellular vestibule of each protomer and influx of water into the protein interior. FRET experiments with SNAP-tagged wild-type and F220C opsin expressed in HEK293 cells are consistent with this conformational difference. We speculate that the unusual mode of dimerization of F220C opsin in the membrane may have physiological consequences.George KhelashviliAnoop Narayana PillaiJoon LeeKalpana PandeyAlexander M. PayneZarek SiegelMichel A. CuendetTylor R. LewisVadim Y. ArshavskyJohannes BroichhagenJoshua LevitzAnant K. MenonNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-20 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q George Khelashvili Anoop Narayana Pillai Joon Lee Kalpana Pandey Alexander M. Payne Zarek Siegel Michel A. Cuendet Tylor R. Lewis Vadim Y. Arshavsky Johannes Broichhagen Joshua Levitz Anant K. Menon Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
description	Abstract Mutations in the G protein-coupled receptor (GPCR) rhodopsin are a common cause of autosomal dominant retinitis pigmentosa, a blinding disease. Rhodopsin self-associates in the membrane, and the purified monomeric apo-protein opsin dimerizes in vitro as it transitions from detergent micelles to reconstitute into a lipid bilayer. We previously reported that the retinitis pigmentosa-linked F220C opsin mutant fails to dimerize in vitro, reconstituting as a monomer. Using fluorescence-based assays and molecular dynamics simulations we now report that whereas wild-type and F220C opsin display distinct dimerization propensities in vitro as previously shown, they both dimerize in the plasma membrane of HEK293 cells. Unexpectedly, molecular dynamics simulations show that F220C opsin forms an energetically favored dimer in the membrane when compared with the wild-type protein. The conformation of the F220C dimer is unique, with transmembrane helices 5 and 6 splayed apart, promoting widening of the intracellular vestibule of each protomer and influx of water into the protein interior. FRET experiments with SNAP-tagged wild-type and F220C opsin expressed in HEK293 cells are consistent with this conformational difference. We speculate that the unusual mode of dimerization of F220C opsin in the membrane may have physiological consequences.
format	article
author	George Khelashvili Anoop Narayana Pillai Joon Lee Kalpana Pandey Alexander M. Payne Zarek Siegel Michel A. Cuendet Tylor R. Lewis Vadim Y. Arshavsky Johannes Broichhagen Joshua Levitz Anant K. Menon
author_facet	George Khelashvili Anoop Narayana Pillai Joon Lee Kalpana Pandey Alexander M. Payne Zarek Siegel Michel A. Cuendet Tylor R. Lewis Vadim Y. Arshavsky Johannes Broichhagen Joshua Levitz Anant K. Menon
author_sort	George Khelashvili
title	Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
title_short	Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
title_full	Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
title_fullStr	Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
title_full_unstemmed	Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin
title_sort	unusual mode of dimerization of retinitis pigmentosa-associated f220c rhodopsin
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/dd6f556860b34a2ea32087d0f5c5847f
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Unusual mode of dimerization of retinitis pigmentosa-associated F220C rhodopsin

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