title: Purification and isolation of choline acetyltransferase from the electric organ of Torpedo marmorata by affinity chromatography.
abstract: Choline acetyltransferase (EC 2.3.1.6) catalyzes the synthesis of the neurotransmitter acetylcholine from acetylcoenzyme A and choline. It has been purified from the electric organ of Torpedo marmorata by a new double-affinity chromatography. Our rapid and specific purification procedure includes affinity chromatography on CoA-Sepharose and then a second affinity chromatography on the enzyme's inhibitor [2-[3-(2-ammonioethoxy)-benzoyl]ethyl]trimethylammonium bromide coupled to Sepharose via a six-carbon spacer arm. The final enzyme preparation has been purified 7300-fold to a specific activity of 73 mumol acetylcholine formed min-1 mg protein-1. The isolated enzyme gave a single band on disc polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The relative molecular mass was determined to be 68,300 +/- 2100.
authors.affiliation: Department of Pharmacology, University of Zurich, Switzerland.
authors.forename: A J
authors.initials: AJ
authors.lastname: Raeber
authors.forename: G
authors.initials: G
authors.lastname: Riggio
authors.forename: P G
authors.initials: PG
authors.lastname: Waser
mesh_terms: D000818:Animals; D002795:Choline O-Acetyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004557:Electric Organ / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008024:Ligands; D008970:Molecular Weight; D014101:Torpedo
chemical_list: D008024:Ligands; D002795:Choline O-Acetyltransferase