Kabishev A A, Patrushev L I
Biokhimiia. 1987 May;52(5):761-71.
A new effective method for purification of rabbit liver N-acetyltransferase to apparent homogeneity has been developed. The method consists in polymin P and ammonium sulfate fractionation, DEAE-Sephacel and Ultragel AcA 44 chromatography and chromatofocusing. In final preparations the enzyme was purified 2500-3000-fold and its specific activity was found to be about 3000-4000 units per mg of protein. During chromatofocusing of enzyme preparations on a middle pressure chromatograph FPLC (Pharmacia, Sweden) a partial separation of acetyltransferase allelic forms from fast and slow-acetylators took place. The supposed allelic acetyltransferase forms differ in some biochemical properties. In particular, the slow acetyltransferase form is much more sensitive towards 0.1 M KCl against the rapid enzyme form. It is assumed that the differences between the catalytic properties of acetyltransferase from rapid and slow acetylators may be explained by differences between their polypeptide primary structures.
已开发出一种将兔肝N - 乙酰转移酶纯化至表观均一性的新有效方法。该方法包括多聚胺P和硫酸铵分级分离、DEAE - 琼脂糖凝胶和Ultragel AcA 44柱色谱以及层析聚焦。在最终制剂中，该酶纯化了2500 - 3000倍，其比活性约为每毫克蛋白质3000 - 4000单位。在使用中压色谱仪FPLC（瑞典Pharmacia公司）对酶制剂进行层析聚焦过程中，快速乙酰化者和慢速乙酰化者的乙酰转移酶等位基因形式发生了部分分离。推测的等位基因乙酰转移酶形式在某些生化特性上有所不同。特别是，慢速乙酰转移酶形式对0.1M KCl的敏感性远高于快速酶形式。据推测，快速和慢速乙酰化者的乙酰转移酶催化特性差异可能由其多肽一级结构的差异来解释。
