Počet záznamů: 1
Plasma membrane cholesterol level and agonist-induced internalization of delta-opioid receptors; colocalization study with intracellular membrane markers of Rab family\n
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SYSNO ASEP 0463372 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Plasma membrane cholesterol level and agonist-induced internalization of delta-opioid receptors; colocalization study with intracellular membrane markers of Rab family
Tvůrce(i) Brejchová, Jana (FGU-C) RID, ORCID
Vošahlíková, Miroslava (FGU-C) RID, ORCID, SAI
Roubalová, Lenka (FGU-C) RID, ORCID, SAI
Parenti, M. (IT)
Mauri, M. (IT)
Chernyavskiy, Oleksandr (FGU-C)
Svoboda, Petr (FGU-C) RID, ORCIDZdroj.dok. Journal of Bioenergetics and Biomembranes - ISSN 0145-479X
Roč. 48, č. 4 (2016), s. 375-396Poč.str. 22 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova cholesterol ; plasma membrane ; delta-opioid receptor ; internalization ; Rab proteins Vědní obor RIV CE - Biochemie CEP GAP207/12/0919 GA ČR - Grantová agentura ČR Institucionální podpora FGU-C - RVO:67985823 UT WOS 000384436600004 EID SCOPUS 84978087082 DOI 10.1007/s10863-016-9667-7 Anotace 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. Pracoviště Fyziologický ústav Kontakt Lucie Trajhanová, lucie.trajhanova@fgu.cas.cz, Tel.: 241 062 400 Rok sběru 2017
Počet záznamů: 1