THE DETERMINATION OF PICOMOLE AMOUNTS OF ACETYLCHOLINE IN MAMMALIAN BRAIN
THE DETERMINATION OF PICOMOLE AMOUNTS OF ACETYLCHOLINE IN MAMMALIAN BRAIN
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哺乳动物脑中乙酰胆碱皮摩尔量的测定
DOI：
10.1111/j.1471-4159.1973.tb12097.x
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发表时间：
1973
影响因子：
4.7
通讯作者：
R. Mccaman
中科院分区：
文献类型：
--
作者：
A. M. Goldberg;R. Mccaman
摘要：在任何基于胆碱转化为产物的测定乙酰胆碱的试验中，直接的问题是去除内源性胆碱。其他已发表的酶测定法已经利用电泳来实现这一目标。在即将描述的实验中，这是通过胆碱激酶对内源性胆碱的酶磷酸化来完成的。一旦这个反应完成，内源性乙酰胆碱同时被水解，然后被[32P]ATP磷酸化。通过ATP的沉淀将标记产物[32P]磷酸胆碱从标记的底物中分离出来，并在离子交换树脂微柱上进一步分离。使用这种方法，可以测量从组织中提取的乙酰胆碱的皮摩尔量。
Abstract— In any assay for the determination of acetylcholine based on the conversion of choline to a product, the immediate problem is the removal of endogenous choline. Other published enzymatic assays have taken advantage of electrophoresis to accomplish this goal. In the assay to be described, this is accomplished by the enzymatic phosphorylation of endogenous choline by choline kinase. Once this reaction is complete, endogenous acetylcholine is simultaneously hydrolysed and then phosphorylated with [32P]ATP. The labelled product [32P]phosphorylcholine is separated from the labelled substrate by precipitation of the ATP and further separation is accomplished on microcolumns of ion exchange resin. Using this methodology, picomole amounts of acetylcholine, derived from tissue, can be measured.