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Kappa but not delta or mu opioid receptors form homodimers at low membrane densities.

ABSTRACT: Opioid receptors (ORs) have been observed as homo- and heterodimers, but it is unclear if the dimers are stable under physiological conditions, and whether monomers or dimers comprise the predominant fraction in a cell. Here, we use three live-cell imaging approaches to assess dimerization of ORs at expression levels that are 10-100 × smaller than in classical biochemical assays. At membrane densities around 25/µm², a split-GFP assay reveals that κOR dimerizes, while µOR and δOR stay monomeric. At receptor densities < 5/µm², single-molecule imaging showed no κOR dimers, supporting the concept that dimer formation depends on receptor membrane density. To directly observe the transition from monomers to dimers, we used a single-molecule assay to assess membrane protein interactions at densities up to 100 × higher than conventional single-molecule imaging. We observe that κOR is monomeric at densities < 10/µm² and forms dimers at densities that are considered physiological. In contrast, µOR and δOR stay monomeric even at the highest densities covered by our approach. The observation of long-lasting co-localization of red and green κOR spots suggests that it is a specific effect based on OR dimerization and not an artefact of coincidental encounters.

SUBMITTER: Cechova K

PROVIDER: S-EPMC8629795 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Kappa but not delta or mu opioid receptors form homodimers at low membrane densities.

Cechova Kristina K Lan Chenyang C Macik Matus M Barthes Nicolas P F NPF Jung Manfred M Ulbrich Maximilian H MH

Cellular and molecular life sciences : CMLS 20211017 23

Opioid receptors (ORs) have been observed as homo- and heterodimers, but it is unclear if the dimers are stable under physiological conditions, and whether monomers or dimers comprise the predominant fraction in a cell. Here, we use three live-cell imaging approaches to assess dimerization of ORs at expression levels that are 10-100 × smaller than in classical biochemical assays. At membrane densities around 25/µm<sup>2</sup>, a split-GFP assay reveals that κOR dimerizes, while µOR and δOR stay ...[more]

PMID: 34657173

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Dataset Information

Kappa but not delta or mu opioid receptors form homodimers at low membrane densities.

Publications

Kappa but not delta or mu opioid receptors form homodimers at low membrane densities.

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