Characterization of curaremimetic neurotoxin binding sites on cellular membrane fragments derived from the rat pheochromocytoma PC12.
Studies were conducted on the properties of 125I-labeled alpha-bungarotoxin binding sites on cellular membrane fragments derived from the PC12 rat pheochromocytoma. Two classes of specific toxin binding sites are present at approximately equal densities (50 fmol/mg of membrane protein) and are characterized by apparent dissociation constants of 3 and 60 nM. Nicotine and d-tubocurarine are among the most potent inhibitors of high-affinity toxin binding. The affinity of high-affinity toxin binding sites for nicotinic cholinergic agonists is reversibly or irreversibly decreased, respectively, on treatment with dithiothreitol or dithiothreitol and N-ethylmaleimide. The nicotinic receptor affinity reagent bromoacetylcholine irreversibly blocks high-affinity toxin binding to PC12 cell membranes that have been treated with dithiothreitol. Two polyclonal antisera raised against the nicotinic acetylcholine receptor from Electrophorus electricus inhibit high-affinity toxin binding. These detailed studies confirm that curaremimetic neurotoxin binding sites on the PC12 cell line are comparable to toxin binding sites from neural tissues and to nicotinic acetylcholine receptors from the periphery. Because toxin binding sites are recognized by anti-nicotinic receptor antibodies, the possibility remains that they are functionally analogous to nicotinic receptors.
D000109:Acetylcholine / Q000378:metabolism; D000310:Adrenal Gland Neoplasms / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002038:Bungarotoxins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D009498:Neurotoxins / Q000378:metabolism*; D009538:Nicotine / Q000378:metabolism; D010673:Pheochromocytoma / Q000378:metabolism*; D051381:Rats; D014101:Torpedo; D014403:Tubocurarine / Q000378:metabolism
D002038:Bungarotoxins; D009498:Neurotoxins; D009538:Nicotine; D000109:Acetylcholine; D004229:Dithiothreitol; D014403:Tubocurarine