Effects of glycosylation inhibitors on human growth hormone receptor in cultured human lymphocytes.
IM-9 cultured human lymphocytes were treated with N-linked glycosylation inhibitors, N-linked oligosaccharide processing inhibitors, or neuraminidase to study the effect of glycosylation modification on human growth hormone binding and molecular weight of surface hGH receptor. One mg/l tunicamycin and 20 mmol/l glucosamine decreased 125I-hGH binding to the cells to 46.3 +/- 2.4% (mean +/- SEM) and 21.9 +/- 0.2% of the controls, respectively. The hGH binding was 33.0 +/- 18.4% of the control value in the cells treated with monensin. The inhibition of binding was due to a decrease in the hGH receptor number without any affinity changes in these cells. Neither 1 mg/l swainsonine nor 100 mg/l castanospermine had any effect on the hGH binding. On the other hand, 125I-hGH binding to neuraminidase-treated cells was significantly enhanced with accompanying affinity changes. When 125I-hGH was cross-linked to IM-9 cells, there were no differences in the molecular weight of hGH receptor complexes (140 K) between untreated cells and cells treated with tunicamycin, glucosamine, monensin, or castanospermine. However, the 128 K hGH-receptor complex appeared in swainsonine-treated cells; this complex was sensitive to endoglycosidase H. These data show that the altered carbohydrate moiety changed hGH binding and the size of surface hGH receptor and suggest that glycosylation of receptor is important for the binding of hGH and for its physiological action.
D000470:Alkaloids / Q000494:pharmacology; D002478:Cells, Cultured; D005944:Glucosamine / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D007212:Indolizines; D008214:Lymphocytes / Q000378:metabolism*; D008985:Monensin / Q000494:pharmacology; D009439:Neuraminidase / Q000494:pharmacology; D011986:Receptors, Somatotropin / Q000187:drug effects*; D017026:Swainsonine; D014415:Tunicamycin / Q000494:pharmacology; D054836:Indolizidines
D000470:Alkaloids; D005944:Glucosamine; D007212:Indolizines; D008985:Monensin; D009439:Neuraminidase; D011986:Receptors, Somatotropin; D017026:Swainsonine; D014415:Tunicamycin; C037806:castanospermine; D054836:Indolizidines