A monoclonal antibody interfering with binding and response of the acetylcholine receptor.
Employing a monoclonal antibody raised against the receptor protein, we have probed the mechanism of ligand interaction of the nicotinic acetylcholine receptor from Torpedo marmorata. Antibody WF6 specifically binds to alpha-subunits of the receptor with a stoichiometry of one molecule per receptor monomer. At saturating concentrations, WF6 blocks half of the binding sites for acetylcholine, all of the binding sites for alpha-neurotoxins, and none of the binding sites for representative cholinergic antagonists (with the exception of alpha-toxins) at the receptor. In the presence of saturating concentrations of antibody WF6, acetylcholine (or its agonists) cannot induce T1+ influx into Torpedo membrane vesicles. Rapid oversaturation of the receptor by agonist also cannot overcome this blockade of channel gating. The observed competition patterns of WF6 and representative cholinergic ligands with the receptor are evidence for separate binding sites for groups of ligands and for a network of allosterically linked effector regions at the receptor. The blockade by saturating concentrations of WF6 of the agonist-induced channel gating supports the conclusion that two molecules of agonist are required to activate the receptor-integral ion channel.
D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001667:Binding, Competitive; D002217:Carbachol / Q000378:metabolism; D003039:Cobra Neurotoxin Proteins / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D007473:Ion Channels / Q000276:immunology; D007700:Kinetics; D051379:Mice; D010277:Parasympathomimetics / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000276:immunology*; D014101:Torpedo / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D003039:Cobra Neurotoxin Proteins; D007473:Ion Channels; D010277:Parasympathomimetics; D011978:Receptors, Nicotinic; C025154:alpha-cobratoxin; D002217:Carbachol; D000109:Acetylcholine