Down-regulation of Gi sub-types by prolonged incubation of adipocytes with an A1 adenosine receptor agonist.
作者： A Green , J L Johnson , G Milligan
DOI: 10.1016/S0021-9258(19)34107-9
关键词: Adenosine 、 Biology 、 Adenosine receptor 、 Pertussis toxin 、 Antiserum 、 Adipocyte 、 Endocrinology 、 Alpha (ethology) 、 Cell culture 、 Internal medicine 、 Agonist
摘要: We have reported previously that prolonged incubation of adipocytes with (-)-N6-phenylisopropyl adenosine (PIA) (an A1 receptor agonist) down-regulates receptors. There was a concomitant decrease in pertussis toxin catalyzed ADP-ribosylation 41-kDa peptide thought to be the alpha-subunit Gi. To determine whether this represents true down-regulation G-protein, and if so which three known forms Gi are down-regulated, we used antipeptide antisera specific for alpha-subunits. Serum SG1 recognizes alpha i1 -2, I1C only i1, I3B i3. Rat were maintained primary culture up 7 days 0-1000 nM PIA. Crude membrane preparations analyzed by Western blots. almost complete loss -3, about 50% i2 from PIA-treated cells. The each i detectable after 24 h 300 PIA maximal 4 days. After days, 3 100 Antiserum BN2 demonstrated approximately G-protein beta-subunits cells treated When incubated then washed remove PIA, -3 returned control levels within 5 CS1 detected normal amounts both 43- 47-kDa Gs conclude alpha-subunits down-regulated along rat adipocytes.
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