Lysosomal degradation of receptor-bound 125I-labeled insulin by rat adipocytes: its characterization and dissociation from the short-term biologic effects of insulin.
In this study we used chloroquine to characterize the interalization and lysosomal degradation of receptor-bound 125I-insulin by rat adipocytes and to determine the role of lysosomal processing of insulin in the short-term biologic effects of the hormone. Chloroquine inhibited the degradation of 125I-insulin bound to adipocytes by both association and disslociation experiments. In the former experiments, chloroquine caused a time- and concentration-dependent increase in specifically bound insulin owing to an increase in intact insulin and a decrease in degradation products, as determined by trichloroacetic acid precipitability and gel chromatographic analysis of material extracted from the cells. In the dissociation experiments, 50 microM chloroquine decreased the rate of degradation by two third, as reflected in the release of degradation  to or degraded by isolated plasma membranes, on the degradation of 125I-insulin by proteases in the incubation medium, or on the endocytotic uptake of receptor-bound insulin. Quantitative electron miroscopy, using monomeric ferritin-insulin, showed 50 microM chloroquine doubled the number of lysosomal structures containing ferritin. These findings are consistent with an inhibition by chloroquine of lysosomal degradation of internalized receptor-bound insulin. Chloroquine, at these same concentrations, had no effect on the ability of insulin to stimulate glucose transport and oxidation or to inhibit epinephrine-stimulated lipolysis. In these studies, we show that lysosomal degradation of internalized receptor-bound insulin is not necessary for insulin to cause short-term biologic effects in the adipocyte.
D000273:Adipose Tissue / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis; D005947:Glucose / Q000378:metabolism; D007328:Insulin / Q000032:analysis / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D008854:Microscopy, Electron; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013395:Sucrose / Q000378:metabolism; D013696:Temperature
D007328:Insulin; D012710:Serum Albumin, Bovine; D013395:Sucrose; D002738:Chloroquine; D011972:Receptor, Insulin; D005947:Glucose