一种用于估算胆碱乙酰转移酶的放射化学方法。
A radiochemical method for the estimation of choline acetyltransferase.
作者信息
Fonnum F
出版信息
Biochem J. 1966 Aug;100(2):479-84. doi: 10.1042/bj1000479.
Abstract
- 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.
摘要
- 已设计出一种用于估算胆碱乙酰转移酶（胆碱乙酰化酶）的放射化学方法，该方法涉及从标记的乙酸盐形成标记的乙酰胆碱。2. [1-(14)C]乙酸盐和辅酶A在非限速浓度的乙酰辅酶A合成酶存在下预孵育，生成[1-(14)C]乙酰辅酶A，然后该物质在乙酰转移酶存在下与胆碱反应，生成[(14)C]乙酰胆碱。3. 反应结束时剩余的任何[(14)C]-乙酰辅酶A通过加入过量的羟胺被破坏，[(14)C]乙酰胆碱通过用四苯基硼酸钠（卡立诺司）沉淀从其他标记化合物中分离出来。4. 洗涤后的沉淀物溶解在乙腈-苄醇中，并通过闪烁计数进行估算。5. 讨论了该方法相对于其他方法的优点。
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