A radiochemical method for the estimation of choline acetyltransferase.
1. A radiochemical method for the estimation of choline acetyltransferase (choline acetylase) has been devised which involves the formation of labelled acetylcholine from labelled acetate. 2. [1-(14)C]Acetate and coenzyme A are pre-incubated in the presence of non-rate-limiting concentrations of acetyl-coenzyme A synthetase to give [1-(14)C]acetyl-coenzyme A, which then reacts with choline in the presence of the acetyltransferase to give [(14)C]acetylcholine. 3. Any [(14)C]-acetyl-coenzyme A remaining at the end of the reaction is destroyed by the addition of excess of hydroxylamine, and [(14)C]acetylcholine is freed from other labelled compounds by precipitation with sodium tetraphenylborate (Kalignost). 4. The washed precipitate is dissolved in acetonitrile-benzyl alcohol and estimated by scintillation counting. 5. Advantages over other methods are discussed.
D000818:Animals; D001717:Birds; D001921:Brain / Q000201:enzymology; D002247:Carbon Isotopes; D002794:Choline; D005459:Fluorides / Q000494:pharmacology; D006168:Guinea Pigs; D011817:Rabbits; D011854:Radiochemistry; D051381:Rats
D002247:Carbon Isotopes; D002794:Choline; D005459:Fluorides