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

  1. 1.
    0463372 - FGÚ 2017 RIV US eng J - Článek v odborném periodiku
    Brejchová, Jana - Vošahlíková, Miroslava - Roubalová, Lenka - Parenti, M. - Mauri, M. - Chernyavskiy, Oleksandr - Svoboda, Petr
    Plasma membrane cholesterol level and agonist-induced internalization of delta-opioid receptors; colocalization study with intracellular membrane markers of Rab family
    .
    Journal of Bioenergetics and Biomembranes. Roč. 48, č. 4 (2016), s. 375-396. ISSN 0145-479X. E-ISSN 1573-6881
    Grant CEP: GA ČR(CZ) GAP207/12/0919
    Institucionální podpora: RVO:67985823
    Klíčová slova: cholesterol * plasma membrane * delta-opioid receptor * internalization * Rab proteins
    Kód oboru RIV: CE - Biochemie
    Impakt faktor: 2.576, rok: 2016

    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.
    Trvalý link: http://hdl.handle.net/11104/0262569

     
     
Počet záznamů: 1  

  Tyto stránky využívají soubory cookies, které usnadňují jejich prohlížení. Další informace o tom jak používáme cookies.