腺苷5'-[β-2P]三磷酸的酶法制备

Enzymic preparation of adenosine 5'-[beta-2P]triphosphate.

作者信息

Leung K L, Yamazaki H

出版信息

Can J Biochem. 1977 Mar;55(3):23-6.

Abstract

A simple method for the preparation of adenosine 5'-[beta-32 P]triphosphate is described. When [32P]orthophosphate was incubated with polyadenylate and phosphoenolpyruvate in the presence of polynucletide phosphorylase (EC 2.7.7.8) and pyruvate kinase (EC 2.7.1.40), up to 75% of 32P radioactivity was recovered in ATP. [32P]ATP was purified to 99.5% radiochemical purity by chromatography on polyethyleneimine-cellulose thin-layer plates. Analysis of hydrolysis products of [32P]ATP with apyrase (EC 3.6.1.5) indicates that 32P in the beta-phosphate position accounts for all 32P label in ATP.

摘要

本文描述了一种制备腺苷5'-[β-32P]三磷酸的简单方法。当在多核苷酸磷酸化酶（EC 2.7.7.8）和丙酮酸激酶（EC 2.7.1.40）存在的情况下，将[32P]正磷酸盐与聚腺苷酸和磷酸烯醇丙酮酸一起温育时，高达75%的32P放射性活度在ATP中回收。通过在聚乙烯亚胺 - 纤维素薄层层析板上进行层析，将[32P]ATP纯化至放射化学纯度为99.5%。用腺苷三磷酸双磷酸酶（EC 3.6.1.5）分析[32P]ATP的水解产物表明，β-磷酸位置的32P占ATP中所有32P标记。