Immunochemical studies of the muscarinic acetylcholine receptor.
Muscarinic receptors have been purified from calf forebrain plasma cell membranes by affinity chromatography on a dexetimide-agarose gel. SDS-PAGE analysis showed a single 70 kDa band. Monoclonal antibodies have been prepared against these affinity purified 70 kDa protein(s). One antibody, M-35, immunoprecipitated up to 80% of digitonin-solubilized muscarinic receptors. M-35 had agonist-like effects on guinea-pig myometrium: it increased the intracellular cyclic GMP content, decreased prostaglandin-induced cyclic AMP accumulation and caused muscle contractions. The two first effects were inhibited by atropine. M-35 was used to visualize muscarinic receptors at the surface of human fibroblastic cells. In the particular cell line used, the receptors have a low affinity for pirenzepine, were negatively coupled to adenylate cyclase and mediated increase in the phosphatidyl-inositol breakdown.
D000109:Acetylcholine / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001285:Atropine / Q000494:pharmacology; D001923:Brain Chemistry; D002417:Cattle; D002460:Cell Line; D002846:Chromatography, Affinity; D006152:Cyclic GMP / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000032:analysis; D006168:Guinea Pigs; D006801:Humans; D010890:Pirenzepine / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D014590:Uterine Contraction
D000911:Antibodies, Monoclonal; D011976:Receptors, Muscarinic; D010890:Pirenzepine; D001285:Atropine; D000262:Adenylyl Cyclases; D006152:Cyclic GMP; D000109:Acetylcholine