Effects of monensin on glycosphingolipid metabolism in cultured human proximal tubular cells.

Effects of monensin, a monovalent cationic ionophore which disrupts Golgi apparatus and its related functions, on glycosphingolipid (GSL) metabolism were investigated in cultured human proximal tubular (PT) cells. Monensin (10(-6) M) stimulated [3H]Gal incorporation into GlcCer, GalCer and LacCer by 8.5-fold and 15-fold, respectively, in PT cells as compared to control. In contrast, [3H]Gal incorporation into GbOse3Cer and GM3 remained unchanged and that into GbOse4Cer was decreased 2-fold as compared to control. GSL measured by HPLC revealed that in cells incubated with monensin, GlcCer, GalCer and LacCer levels were increased 1.6-fold and 7-fold, respectively, whereas GbOse3Cer and GbOse4Cer levels were decreased several folds. Cells incubated with monensin contained 2.5- to 3-fold higher activity of alpha-galactosidase, beta-galactosidase and beta-glucosidase than control, whereas the activity of UDP-gal: glucosylceramide galactosyltransferase (beta-GalT-2) was 8-fold lower than control cells. Cells incubated with monensin took up and degraded one-half as much 125I-LDL as that of control cells. In control cells, exogenously derived [3H]LacCer on LDL was rapidly taken up and catabolized to monoglycosylceramide, or it was used for the endogenous synthesis of globotriosylceramide (trihexosylceramide), globotetraosylceramide (tetrahexosylceramide) and a ganglioside, GM3. In contrast, cells incubated with monensin accumulated most of the [3H]LacCer-LDL. Exogenously derived [3H]LacCer on LDL was catabolized to GlcCer, but was not utilized, for the synthesis of globotriosylceramide, globotetraosylceramide and GM3 in cells incubated with monensin.(ABSTRACT TRUNCATED AT 250 WORDS)