title: Affinity purification of nicotinic acetylcholine receptor from rat brain.
abstract: Using chromatography on DE-52 and acetylcholine-Affi-Gel columns, nicotinic acetylcholine receptor was purified to approximately 10,000 fold from Lubrol extract of rat brain with a recovery of 15%. The purified preparation contained no cholinesterase activity. alpha-Bungarotoxin did not inhibit [3H]acetylcholine binding to the purified preparation. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed 4 major protein bands with apparent molecular weights of 53,000, 67,000, 80,000 and 108,000. When nicotinic acetylcholine receptor was eluted with either carbachol or nicotine from the affinity column, these major bands were found on SDS-PAGE gels. Immunoblot analysis showed that the Mr 80,000 protein was an acetylcholine-binding subunit and that the Mr 48,000 protein, a minor band on SDS-PAGE gel, was a structural subunit.
authors.affiliation: Department of Pharmacology, Nara Medical University, Kashihara, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Nakayama
authors.forename: M
authors.initials: M
authors.lastname: Shirase
authors.forename: T
authors.initials: T
authors.lastname: Nakashima
authors.forename: Y
authors.initials: Y
authors.lastname: Kurogochi
authors.forename: J M
authors.initials: JM
authors.lastname: Lindstrom
mesh_terms: D000109:Acetylcholine / Q000031:analogs & derivatives*; D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000378:metabolism*; D002846:Chromatography, Affinity; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D011978:Receptors, Nicotinic / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D011978:Receptors, Nicotinic; D000109:Acetylcholine