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Kappa but not delta or mu opioid receptors form homodimers at low membrane densities
K. Cechova, C. Lan, M. Macik, NPF. Barthes, M. Jung, MH. Ulbrich
Language English Country Switzerland
Document type Journal Article
Grant support
UL 312/6-1
deutsche forschungsgemeinschaft
RTG 2202
deutsche forschungsgemeinschaft
EXC 294
deutsche forschungsgemeinschaft
CRC 992
deutsche forschungsgemeinschaft
EXC-2189-Project ID: 390939984
deutsche forschungsgemeinschaft
GA17-05903S
grantová agentura české republiky
SVV260427/2020
univerzita karlova v praze
FM/a/2017-2-072
univerzita karlova v praze
SVV260427/2018
univerzita karlova v praze
NLK
PubMed Central
from 1997
ProQuest Central
from 1997-01-01 to 1 year ago
Medline Complete (EBSCOhost)
from 2000-01-01 to 1 year ago
Health & Medicine (ProQuest)
from 1997-01-01 to 1 year ago
- MeSH
- Single-Cell Analysis methods MeSH
- Cell Membrane metabolism MeSH
- Protein Conformation MeSH
- Rats MeSH
- Protein Multimerization MeSH
- Mice MeSH
- Receptors, Opioid, delta chemistry metabolism MeSH
- Receptors, Opioid, mu chemistry metabolism MeSH
- Single Molecule Imaging methods MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
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.
BIOSS Centre for Biological Signalling Studies University of Freiburg Freiburg Germany
CIBSS Centre for Integrative Biological Signalling Studies University of Freiburg Freiburg Germany
Department of Biochemistry Faculty of Science Charles University Prague Czech Republic
Department of Biomathematics Institute of Physiology Czech Academy of Sciences Prague Czech Republic
Faculty of Biology University of Freiburg Freiburg Germany
Institute of Internal Medicine 4 Medical Center of the University of Freiburg Freiburg Germany
Institute of Pharmaceutical Sciences University of Freiburg Freiburg Germany
References provided by Crossref.org
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- $a 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.
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