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Cryo-EM structure of the native rhodopsin dimer in nanodiscs

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dc.abstract.enImaging of rod photoreceptor outer-segment disc membranes by atomic force microscopy and cryo-electron tomography has revealed that the visual pigment rhodopsin, a prototypical classAGprotein-coupled receptor (GPCR), can organize as rows of dimers. GPCR dimerization and oligomerization offer possibilities for allosteric regulation of GPCR activity, but the detailed structures and mechanism remain elusive. In this investigation, we made use of the high rhodopsin density in the native disc membranes and of a bifunctional cross-linker that preserves the native rhodopsin arrangement by covalently tethering rhodopsins via Lys residue side chains. We purified crosslinked rhodopsin dimers and reconstituted them into nanodiscs for cryo-EM analysis. We present cryo-EM structures of the cross-linked rhodopsin dimer as well as a rhodopsin dimer reconstituted into nanodiscs from purified monomers. We demonstrate the presence of a preferential 2-fold symmetrical dimerization interface mediated by transmembrane helix 1 and the cytoplasmic helix 8 of rhodopsin. We confirmed this dimer interface by double electron- electron resonance measurements of spin-labeled rhodopsin. We propose that this interface and the arrangement of two protomers is a prerequisite for the formation of the observed rows of dimers. We anticipate that the approach outlined here could be extended to other GPCRs or membrane receptors to better understand specific receptor dimerization mechanisms.
dc.affiliationUniwersytet Warszawski
dc.contributor.authorPlitzko, Jürgen M.
dc.contributor.authorBaumeister, Wolfgang
dc.contributor.authorFilipek, Sławomir
dc.contributor.authorErnst, Oliver P.
dc.contributor.authorPalczewski, Krzysztof
dc.contributor.authorGulati, Sahil
dc.contributor.authorEps, Ned Van
dc.contributor.authorZhang, Jianye
dc.contributor.authorZhao, Dorothy Yanling
dc.contributor.authorPöge, Matthias
dc.contributor.authorMorizumi, Takefumi
dc.contributor.authorMiszta, Przemysław
dc.contributor.authorMahamid, Julia
dc.date.accessioned2024-01-24T20:55:38Z
dc.date.available2024-01-24T20:55:38Z
dc.date.copyright2019-08-09
dc.date.issued2019
dc.description.accesstimeAT_PUBLICATION
dc.description.financeNie dotyczy
dc.description.number39
dc.description.versionFINAL_PUBLISHED
dc.description.volume294
dc.identifier.doi10.1074/JBC.RA119.010089
dc.identifier.issn0021-9258
dc.identifier.urihttps://repozytorium.uw.edu.pl//handle/item/103982
dc.identifier.weblinkhttps://api.elsevier.com/content/article/PII:S0021925820349838?httpAccept=text/xml
dc.languageeng
dc.pbn.affiliationchemical sciences
dc.relation.ispartofJournal of Biological Chemistry
dc.relation.pages14215-14230
dc.rightsCC-BY
dc.sciencecloudnosend
dc.titleCryo-EM structure of the native rhodopsin dimer in nanodiscs
dc.typeJournalArticle
dspace.entity.typePublication