Covalent cross-linking of prostaglandin E receptor from bovine adrenal medulla with a pertussis toxin-insensitive guanine nucleotide-binding protein.

摘要: Abstract Prostaglandin E2 (PGE2) specifically bound to 100,000 X g pellet prepared from bovine adrenal medulla, and [3H]PGE2-bound proteins were solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid. The dissociation of [3H]PGE2 the was enhanced by GTP. [3H]PGE2-specifically adsorbed onto a wheat germ agglutinin column GTP treatment decreased amount retained on column. When cross-linked in membrane dithiobis(succinimidyl propionate) solubilized, no longer dissociated treatment, suggesting that cross-linking produced stable high-affinity complex PGE receptor GTP-binding protein. Covalent attested adsorption propionate)-treated GTP-Sepharose, co-elution [35S]guanosine 5'-O-(3-thiotriphosphate) binding activity immunoreactivities alpha o beta subunits eluted as an apparently single radioactive peak at position Mr = 200,000 gel filtration. These results have demonstrated is glycoprotein approximate 110,000, assuming protein 90,000. PGE2 neither activated nor inhibited adenylate cyclase activity, pertussis toxin (islet-activating protein) did not affect its sensitivity. suggest may be functionally associated toxin-insensitive coupled system medulla.