Degradation of insulin receptors by hepatoma cells: insulin-induced down-regulation results from an increase in the rate of basal receptor degradation.
The degradation of insulin receptors was studied in cultured Zajdela hepatoma cells (ZHC). Receptor distribution within the cell was evaluated by estimating: i) surface receptor level on entire cells, ii) total cell receptors solubilized by Triton from cell membranes and iii) intracellular receptors solubilized from cells whose surface receptors had been inactivated with trypsin. In the absence of insulin, 80-90% of the insulin binding sites were located on the cell surface. When insulin was added, a rapid decrease of surface receptors was observed. After 2 h, their level was reduced nearly by half; this reduction was accounted for by an actual receptor loss from the cell without an increase in the intracellular pool. These results indicate that insulin enhanced the rate of receptor degradation within the cell. Basal receptor inactivation was studied by using tunicamycin which inhibits new receptor synthesis. The surface receptor number was decreased with a half-life of 7 h, while the level of internal sites remained unchanged. Both basal and insulin-activated receptor degradation were markedly slowed down by chloroquine or dansylcadaverine, indicating the importance of endocytic pathways in this process. Similarly, when de novo protein glycosylation was inhibited for 24 h by tunicamycin, both basal and insulin-activated receptor inactivation were precluded.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D051381:Rats; D011972:Receptor, Insulin / Q000187:drug effects / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology
D007328:Insulin; D014415:Tunicamycin; D002738:Chloroquine; D011972:Receptor, Insulin; D014357:Trypsin; C008542:monodansylcadaverine; D002103:Cadaverine