Szyfter K, Hemminki K, Crane A E, Watson W P
Institute of Occupational Health, Helsinki, Finland.
Chem Biol Interact. 1991;80(1):99-107. doi: 10.1016/0009-2797(91)90034-5.
1,N6-ethenodeoxyadenosine-, 1,N2-ethenodeoxyguanosine- and 3,N4-ethenodeoxycytidine-3'-monophosphates were labeled by [gamma-32P] ATP using T4 polynucleotide kinase in conditions commonly used for the 32P-postlabeling assay. Kinetic studies showed that the reaction is fast reaching a plateau after 15-30 min. The efficiency of phosphorylation, as studied by substrate-product concentration dependency, was between 50-100% at the lower substrate concentrations. The adducts are labeled efficiently at sub-femtomole levels. All the adducts were sensitive to the 3'-dephosphorylation by P1 nuclease although the guanine derivative appeared to be more resistant than the two other adducts.
使用T4多核苷酸激酶，在常用于32P后标记分析的条件下，用[γ-32P]ATP对1,N6-乙烯基脱氧腺苷-3'-单磷酸、1,N2-乙烯基脱氧鸟苷-3'-单磷酸和3,N4-乙烯基脱氧胞苷-3'-单磷酸进行标记。动力学研究表明，该反应速度很快，15 - 30分钟后达到平稳状态。通过底物 - 产物浓度依赖性研究发现，在较低底物浓度下，磷酸化效率在50 - 100%之间。这些加合物在亚飞摩尔水平就能被有效标记。所有加合物对P1核酸酶的3'-去磷酸化反应敏感，不过鸟嘌呤衍生物似乎比其他两种加合物更具抗性。
