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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F21%3A00549802" target="_blank" >RIV/67985823:_____/21:00549802 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216208:11310/21:10441796

Výsledek na webu

<a href="https://doi.org/10.1007/s00018-021-03963-y" target="_blank" >https://doi.org/10.1007/s00018-021-03963-y</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s00018-021-03963-y" target="_blank" >10.1007/s00018-021-03963-y</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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 x smaller than in classical biochemical assays. At membrane densities around 25/mu m(2), a split-GFP assay reveals that kappa OR dimerizes, while mu OR and delta OR stay monomeric. At receptor densities < 5/mu m(2), single-molecule imaging showed no kappa OR 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 x higher than conventional single-molecule imaging. We observe that kappa OR is monomeric at densities < 10/mu m(2) and forms dimers at densities that are considered physiological. In contrast, mu OR and delta OR stay monomeric even at the highest densities covered by our approach. The observation of long-lasting co-localization of red and green kappa OR spots suggests that it is a specific effect based on OR dimerization and not an artefact of coincidental encounters.

Název v anglickém jazyce

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

Popis výsledku anglicky

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 x smaller than in classical biochemical assays. At membrane densities around 25/mu m(2), a split-GFP assay reveals that kappa OR dimerizes, while mu OR and delta OR stay monomeric. At receptor densities < 5/mu m(2), single-molecule imaging showed no kappa OR 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 x higher than conventional single-molecule imaging. We observe that kappa OR is monomeric at densities < 10/mu m(2) and forms dimers at densities that are considered physiological. In contrast, mu OR and delta OR stay monomeric even at the highest densities covered by our approach. The observation of long-lasting co-localization of red and green kappa OR spots suggests that it is a specific effect based on OR dimerization and not an artefact of coincidental encounters.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/GA17-05903S" target="_blank" >GA17-05903S: Změny mobility a funkce receptoru pro opioidy typu δ vyvolané akutní a chronickou deplecí cholesterolu v plazmatických membránách živých buněk</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Cellular and Molecular Life Sciences

ISSN

1420-682X

e-ISSN

1420-9071

Svazek periodika

78

Číslo periodika v rámci svazku

23

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

12

Strana od-do

7557-7568

Kód UT WoS článku

000707846600001

EID výsledku v databázi Scopus

2-s2.0-85117590664