Kavunenko A P, Kalacheva T N, Tikhomirova-Sidorova N S
Biokhimiia. 1979 Aug;44(8):1381-91.
In order to determine the relative activity of pyrimidine nucleoside-2',3'-cyclophosphates as donors and nucleosides as acceptors of phosphate in the reaction of the internucleotide bond formation catalyzed by RNAase A (EC 3.4.1.22), a comparative synthesis of dinucleoside monophosphates UpU, UpC, CpU and CpC at three different enzyme concentrations (20, 40 and 70 mkg/ml) and two temperatures (0 degrees and -15 degrees) was carried out. The conversion rate of donor (U greater than p and C greater than p) during the synthesis and in the competitive reaction of hydrolysis strongly depends on the type of acceptor activity as compared to uridine. Based on the data of synthesis and simultaneous hydrolysis of U greater than p and C greater than p it may be concluded that in the both cases the latter donor is more reactive. The approaches to the determination of the substrate activity of the donors and acceptors for the evaluation of optimal conditions of the dinucleoside monophosphate synthesis depending on the donor--acceptor combination are discussed.
为了确定嘧啶核苷 - 2',3'-环磷酸酯作为供体以及核苷作为受体在核糖核酸酶A(EC 3.4.1.22)催化的核苷酸间键形成反应中磷酸基团的相对活性,在三种不同的酶浓度(20、40和70微克/毫升)以及两个温度(0℃和 - 15℃)下进行了二核苷单磷酸UpU、UpC、CpU和CpC的比较合成。与尿苷相比,供体(U>p和C>p)在合成过程以及水解竞争反应中的转化率强烈依赖于受体活性的类型。基于U>p和C>p的合成及同时水解的数据可以得出结论,在这两种情况下,后一种供体更具反应活性。讨论了根据供体 - 受体组合来确定供体和受体的底物活性以评估二核苷单磷酸合成最佳条件的方法。