Effect of tunicamycin on functions of PGE1 receptors from mouse mastocytoma P-815 cells.
Prostaglandin E1 (PGE1) receptors from mouse mastocytoma P-815 cells were found to bind to a wheat germ agglutinin (WGA)-Agarose column, suggesting that the receptors are glycoproteins. To further elucidate the role of carbohydrate moieties in the PGE1 receptors for their binding activity to ligand, the P-815 cells were treated with tunicamycin, swainsonine or monensin. Tunicamycin, an inhibitor of N-glycosylation, dose- and time-dependently inhibited the binding of PGE1 to mastocytoma P-815 cells. Neither swainsonine, an inhibitor of Golgi mannosidase II, nor monensin, an inhibitor of processing beyond the high mannose stage, altered PGE1 binding properties of the cells. The inhibition of PGE1 binding by tunicamycin was observed when incorporation of [3H]glucosamine into macromolecules was inhibited. The inhibitory effect was not on their affinity but on their number of binding sites. Subcellular distributions of [3H]PGE1-binding activity showed that decreases in the binding activity by tunicamycin were highest in plasma membrane fractions. Treatment of membranes with various endo- and exoglycosidases did not affect PGE1 binding. PGE1-stimulated cyclic AMP accumulation in the cells was also inhibited by tunicamycin. These results suggest that PGE1 receptors of mastocytoma P-815 cells are glycoproteins and that inhibition of N-glycosylation of PGE1 receptors by tunicamycin results in the arrest of the translocation of newly synthesized receptors to the surface of mastocytoma P-815 cells.
Yatsunami
K
K
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Fujisawa
J
J
Hashimoto
H
H
Kimura
K
K
Takahashi
S
S
Ichikawa
A
A
D000470:Alkaloids / Q000494:pharmacology; D000527:Alprostadil / Q000378:metabolism*; D000818:Animals; D002853:Chromatography, Liquid; D000242:Cyclic AMP / Q000378:metabolism; D006031:Glycosylation; D012515:Mast-Cell Sarcoma / Q000378:metabolism*; D051379:Mice; D008985:Monensin / Q000494:pharmacology; D011982:Receptors, Prostaglandin / Q000187:drug effects*; D017026:Swainsonine; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology*
D000470:Alkaloids; D011982:Receptors, Prostaglandin; D014415:Tunicamycin; D008985:Monensin; D000242:Cyclic AMP; D000527:Alprostadil; D017026:Swainsonine