title: Purification of a nicotinic acetylcholine receptor from rat brain by affinity chromatography directed at the acetylcholine binding site.
abstract: We have purified a nicotinic acetylcholine receptor from rat brain by use of an acetylcholine affinity resin commonly employed for the purification of nicotinic acetylcholine receptor from electric tissue. Receptor, specifically eluted with nicotine, bound (-)-[3H]nicotine with a dissociation constant of approximately 21 nM. Binding was inhibited by carbamylcholine but not by alpha-bungarotoxin. Polyacrylamide gel electrophoresis yielded two protein bands, of apparent mol. wts. 80,400 and 52,400. These results provide independent confirmation of the subunit size and composition reported for rat brain nicotinic receptor isolated by immunoaffinity methods and demonstrate a method of purification that can be performed with commercially available reagents.
authors.affiliation: Division of Neuropathology, Columbia University, New York, NY.
authors.forename: A J
authors.initials: AJ
authors.lastname: Dwork
authors.forename: J T
authors.initials: JT
authors.lastname: Desmond
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008970:Molecular Weight; D009538:Nicotine / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011978:Receptors, Nicotinic / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D011978:Receptors, Nicotinic; D009538:Nicotine; D000109:Acetylcholine