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胆碱乙酰转移酶(choline-O-acetyhran-sferase,ChAT,EC 2.3.1.6)活性,目前文献上最常用放射化学法测定。其基本原理是~(14)C或~3H标记的底物——乙酰辅酶A(Ac-CoA)在ChAT的催化下,把标记乙酰基转移到胆碱(Ch)上,生成标记的乙酰胆碱(AOh)。[~(14)C]Ac-CoA+Ch=[~(14)C]ACh+CoA 通过适当的方法,把生成的标记产物和底物分离,测定产物放射性,即可判断酶活性。因此如何特异地、有效地分离生成物和底物,是方法的关键。文献上多采用离子交换法、选择性沉淀法、电泳法、有机提取法和多种方法联合应用。我们采用
The activity of choline-O-acetyhran-sferase (ChAT, EC 2.3.1.6) is currently the most commonly used radiochemical assay. Its basic principle is ~ (14) C or ~ 3H labeled substrate - acetyl CoA (Ac-CoA) under the catalysis of ChAT, transfer the labeled acetyl to choline (Ch) to generate labeled acetylcholine AOh). [~ (14) C] Ac-CoA + Ch = [~ (14) C] ACh + CoA The enzyme activity can be judged by measuring the radioactivity of the product by separating the labeled product from the substrate by a suitable method. Therefore, how to separate the products and substrates specifically and effectively is the key of the method. Literature more use of ion exchange, selective precipitation, electrophoresis, organic extraction and a variety of methods used in combination. We use