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  Cryo-EM structure of the native rhodopsin dimer in nanodiscs

Zhao, D. Y., Pöge, M., Morizumi, T., Gulati, S., Van Eps, N., Zhang, J., et al. (2019). Cryo-EM structure of the native rhodopsin dimer in nanodiscs. JOURNAL OF BIOLOGICAL CHEMISTRY, 294(39), 14215-14230. doi:10.1074/jbc.RA119.010089.

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 Creators:
Zhao, Dorothy Yanling1, Author
Pöge, Matthias2, Author              
Morizumi, Takefumi1, Author
Gulati, Sahil1, Author
Van Eps, Ned1, Author
Zhang, Jianye1, Author
Miszta, Przemyslaw1, Author
Filipek, Slawomir1, Author
Mahamid, Julia1, Author
Plitzko, Jürgen M.2, Author              
Baumeister, Wolfgang2, Author              
Ernst, Oliver P.1, Author
Palczewski, Krzysztof1, Author
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1external, ou_persistent22              
2Baumeister, Wolfgang / Molecular Structural Biology, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565142              

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Free keywords: PROTEIN-COUPLED RECEPTOR; CRYSTAL-STRUCTURE; OPIOID RECEPTOR; C5A RECEPTOR; DIMERIZATION; OLIGOMERIZATION; CELLS; OPSIN; ORGANIZATION; ARRESTING protein-coupled receptor (GPCR); receptor; rhodopsin; retinoid-binding protein; retina; cryo-electron microscopy (cryo-EM); rod outer segment; transmembrane helix 1 (TM1); helix 8 (H8); cell signaling; double electron-electron resonance (DEER); rhodopsin dimerization; transducin;
 Abstract: Imaging of rod photoreceptor outer-segment disc membranes by atomic force microscopy and cryo-electron tomography has revealed that the visual pigment rhodopsin, a prototypical class A G protein?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 cross-linked 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.

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Language(s): eng - English
 Dates: 2019
 Publication Status: Published in print
 Pages: 16
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: ISI: 000499248200006
DOI: 10.1074/jbc.RA119.010089
 Degree: -

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Title: JOURNAL OF BIOLOGICAL CHEMISTRY
Source Genre: Journal
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Publ. Info: 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA : AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
Pages: - Volume / Issue: 294 (39) Sequence Number: - Start / End Page: 14215 - 14230 Identifier: ISSN: 0021-9258