Plasma membrane cholesterol level and agonist-induced internalization of delta-opioid receptors; colocalization study with intracellular membrane markers of Rab familyn

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F67985823%3A_____%2F16%3A00463372" target="_blank" >RIV/67985823:_____/16:00463372 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1007/s10863-016-9667-7" target="_blank" >http://dx.doi.org/10.1007/s10863-016-9667-7</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s10863-016-9667-7" target="_blank" >10.1007/s10863-016-9667-7</a>

Result language

angličtina

Original language name

Plasma membrane cholesterol level and agonist-induced internalization of delta-opioid receptors; colocalization study with intracellular membrane markers of Rab familyn

Original language description

Decrease of cholesterol level in plasma membrane of living HEK293 cells transiently expressing FLAG-delta-OR by beta-cyclodextrin (beta-CDX) resulted in a slight internalization of delta-OR. Massive internalization of delta-OR induced by specific agonist DADLE was diminished in cholesterol-depleted cells. These results suggest that agonist-induced internalization of delta-OR, which has been traditionally attributed exclusively to clathrin-mediated pathway, proceeds at least partially via membrane domains. Identification of internalized pools of FLAG-delta-OR by colocalization studies with proteins of Rab family indicated the decreased presence of receptors in early endosomes (Rab5), late endosomes and lysosomes (Rab7) and fast recycling vesicles (Rab4). Slow type of recycling (Rab11) was unchanged by cholesterol depletion. As expected, agonist-induced internalization of oxytocin receptors was totally suppressed in beta-CDX-treated cells. Determination of average fluorescence lifetime of TMA-DPH, the polar derivative of hydrophobic membrane probe diphenylhexatriene, in live cells by FLIM indicated a significant alteration of the overall PM structure which may be interpreted as an increased "water-accessible space" within PM area. Data obtained by studies of HEK293 cells transiently expressing FLAG-delta-OR by "antibody feeding" method were extended by analysis of the effect of cholesterol depletion on distribution of FLAG-delta-OR in sucrose density gradients prepared from HEK293 cells stably expressing FLAG-delta-OR. Major part of FLAG-delta-OR was co-localized with plasma membrane marker Na,K-ATPase and beta-CDX treatment resulted in shift of PM fragments containing both FLAG-delta-OR and Na,K-ATPase to higher density. Thus, the decrease in content of the major lipid constituent of PM resulted in increased density of resulting PM fragments.

Czech name

—

Czech description

—

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

—

Project

<a href="/en/project/GAP207%2F12%2F0919" target="_blank" >GAP207/12/0919: The role of hydrophobic plasma membrane interior in DOR regulation of trimeric G protein activity</a><br>

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2016

Confidentiality

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

Name of the periodical

Journal of Bioenergetics and Biomembranes

ISSN

0145-479X

e-ISSN

—

Volume of the periodical

48

Issue of the periodical within the volume

4

Country of publishing house

US - UNITED STATES

Number of pages

22

Pages from-to

375-396

UT code for WoS article

000384436600004

EID of the result in the Scopus database

2-s2.0-84978087082