title: Effects of monoclonal antibodies on the function of acetylcholine receptors purified from Torpedo californica and reconstituted into vesicles.
abstract: We tested the effects of 62 monoclonal antibodies (mAbs) to acetylcholine receptors from Torpedo californica on the function of receptor reconstituted into lipid vesicles. Two of these mAbs, mAbs 148 and 168, inhibited carbamylcholine-induced 22Na+ uptake into vesicles. The rate of 125I-labeled alpha-bungarotoxin (125I-alpha BGT) binding to the reconstituted liposomes was also reduced, although 125I-alpha BGT binding at equilibrium was not affected. Agonist-induced desensitization of the receptor was also affected by these mAbs. mAb 148 binds to the beta subunit of receptor, and mAb 168 binds to the gamma subunit. Both mAbs bind to the cytoplasmic surface of the receptor; correspondingly, both block function when added before reconstitution, and both were found to have no effect on function when added to preformed vesicles. Their effects were not due to interference with the reconstitution process. Both mAbs were capable of cross-linking receptors. In contrast to the bivalent mAbs, monovalent Fab fragments of these two mAbs had little effect on receptor function, which suggests that the effects of the bivalent mAbs resulted primarily from cross-linking receptors.
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D002038:Bungarotoxins / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D004557:Electric Organ / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments; D007700:Kinetics; D051379:Mice; D010954:Plasmacytoma / Q000276:immunology; D051381:Rats; D011950:Receptors, Cholinergic / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014101:Torpedo
chemical_list: D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D007140:Immunoglobulin Fab Fragments; D011950:Receptors, Cholinergic; D002217:Carbachol