Doi S, Tanigawa Y, Tsuchiya M, Shimoyama M
Department of Biochemistry, Shimane Medical University, Izumo, Japan.
Biochim Biophys Acta. 1993 Mar 5;1162(1-2):115-20. doi: 10.1016/0167-4838(93)90136-f.
We investigated the effects of acid treatment (pH 3.5) on the activities of arginine-specific ADP-ribosyltransferase (mono(ADP-ribosyl)transferase) (EC 2.4.2.31) and poly(ADP-ribose) synthetase (EC 2.4.2.30) purified from chicken liver, and we observed that the former enzyme retained completely its activity while there was no evidence for activity of the latter enzyme. Kinetic parameters, including Km values for NAD and whole histones in the reaction catalyzed by acid-treated ADP-ribosyltransferase, were the same as those in the reaction catalyzed by non-treated enzyme. The stereospecificities of the reaction forming ADP-ribosylarginine by acid-treated and non-treated ADP-ribosyltransferases were indistinguishable. We made use of this acid treatment to determine ADP-ribosyltransferase activity in chicken and chick-embryo liver extracts, with a filter assay. The enzyme activities (means +/- S.D. for three separate experiments) of 1-year-old chicken and 13-day-old chick embryo livers were 372.7 +/- 80.20 and 110.6 +/- 27.22 nmol ADP-ribose/g wet liver per h, respectively. This acid treatment is useful for filter assay with labelled NAD of arginine-specific ADP-ribosyltransferase in the fraction containing poly(ADP-ribose) synthetase.
我们研究了酸处理（pH 3.5）对从鸡肝中纯化的精氨酸特异性ADP - 核糖基转移酶（单（ADP - 核糖基）转移酶）（EC 2.4.2.31）和聚（ADP - 核糖）合成酶（EC 2.4.2.30）活性的影响，并且我们观察到前一种酶完全保留了其活性，而后一种酶则没有活性的证据。在酸处理的ADP - 核糖基转移酶催化的反应中，包括NAD和全组蛋白的Km值在内的动力学参数与未处理酶催化的反应中的参数相同。酸处理和未处理的ADP - 核糖基转移酶形成ADP - 核糖基精氨酸反应的立体特异性无法区分。我们利用这种酸处理通过滤膜分析法测定鸡和鸡胚肝提取物中的ADP - 核糖基转移酶活性。1岁鸡和13日龄鸡胚肝的酶活性（三个独立实验的平均值±标准差）分别为每小时每克湿肝372.7±80.20和110.6±27.22 nmol ADP - 核糖。这种酸处理对于在含有聚（ADP - 核糖）合成酶的级分中用标记的NAD对精氨酸特异性ADP - 核糖基转移酶进行滤膜分析是有用的。
