title: Studies on the turnover of glucocerebrosidase in cultured rat peritoneal macrophages and normal human fibroblasts.
abstract: The kinetics of glucocerebrosidase synthesis and degradation in rat peritoneal macrophages and in human fibroblasts have been studied using conduritol B epoxide (CBE), an irreversible and specific inhibitor of mammalian glucocerebrosidase. In cultured fibroblasts, higher concentrations of CBE and/or longer times were required for inhibition of glucocerebrosidase than were necessary for inhibition of the macrophage enzyme. However, inhibition of activity in cell extracts from both cell types showed identical time and concentration dependence. After the removal of CBE from cultures, enzyme activity returned to normal with a half-time of 48 h for macrophages and 40 h for fibroblasts. The reappearance of enzyme activity was prevented by an inhibitor of protein synthesis. Both the rate of synthesis and degradation of glucocerebrosidase enzyme protein were independent of the presence of CBE. The calculated rate of degradation of glucocerebrosidase was confirmed using metabolically labelled enzyme in cell cultures. The rate of synthesis for macrophages is 1.8 ng enzyme h-1 mg cell protein-1 and the rate of degradation is 1.4% h-1 (0.014 h-1). These values were 2.0 ng h-1 mg-1 and 0.018 h-1 for fibroblasts.
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D011232:Chemical Precipitation; D005347:Fibroblasts / Q000201:enzymology*; D005959:Glucosidases / Q000378:metabolism*; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007120:Immunochemistry; D007294:Inositol / Q000031:analogs & derivatives / Q000494:pharmacology; D007700:Kinetics; D008264:Macrophages / Q000378:metabolism*; D010529:Peritoneal Cavity; D051381:Rats
chemical_list: D007294:Inositol; D005959:Glucosidases; D005962:Glucosylceramidase; C011423:conduritol epoxide