Inhibition by bacitracin of rat adipocyte plasma membrane degradation of 125I-insulin is associated with an increase in plasma membrane bound insulin and a potentiation of glucose oxidation by adipocytes.
The present study demonstrated that at physiological concentrations of insulin bacitracin inhibited the degradation of specifically bound insulin by enzymes located in the rat adipocyte plasma membrane. Bacitracin increased the amount of intact insulin specifically bound to the plasma membrane and potentiated the stimulation of adipocyte glucose oxidation by submaximal concentrations of the hormone. In contrast to agents such as chloroquine, which inhibit lysosomal degradation of internalized insulin, bacitracin was shown by two approaches to inhibit a degradative process localized to the adipocyte plasma membrane. Cyanide and 2,4-dinitrophenol, agents which inhibit energy requiring endocytosis, had no effect on the bacitracin inhibition of cellular degradation of 125I-insulin. Bacitracin directly inhibited 125I-insulin degradation by isolated plasma membranes at similar concentrations and to a similar extent as found with cells. The degradative process inhibited by bacitracin accounted for the majority of cellular degradation of the hormone. The increased 125I-insulin bound to adipocytes was shown to be intact by gel chromatographic analysis and was localized to the plasma membrane by direct and indirect approaches. Bacitracin increased 125I-insulin specifically bound to isolated plasma membranes as early as 2 min. The 125I-insulin bound to adipocytes in the presence of bacitracin was completely dissociable by the addition of 8 microM unlabeled insulin whereas a significant portion of 125I-insulin bound to chloroquine-treated cells could not be dissociated. Bacitracin slowed dissociation of 125I-insulin from the cells. Bacitracin increased the 125I-insulin binding to cells in the presence and absence of cyanide and 2,4-dinitrophenol. Bacitracin potentiated the stimulation of adipocyte glucose oxidation at submaximal concentrations of insulin.
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D001414:Bacitracin / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects; D007328:Insulin / Q000031:analogs & derivatives* / Q000378:metabolism; D007700:Kinetics; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D011972:Receptor, Insulin / Q000187:drug effects / Q000378:metabolism*
D007328:Insulin; C008147:insulin, iodo-; D001414:Bacitracin; D011972:Receptor, Insulin; D005947:Glucose