A Simple and Sensitive Ribonucleotide Reductase Assay
作者: Jong A.Y.Yu K.Zhou B.Frgala T.Reynolds C.P.Yen Y.
刊名: Journal of Biomedical Science, 1998, Vol.5 (1), pp.62-68
来源数据库: Karger
DOI: 10.1159/000025313
关键词: Ribonucleotide reductaseEnzyme assay
原始语种摘要: Ribonucleotide reductase (RR) is a key regulatory enzyme in the DNA synthesis pathway and is the target of the cancer chemotherapeutic agent hydroxyurea. The study of RR is significantly hindered by the tedious and labor-intensive nature of enzymatic assay. In this report, we present a novel RR assay in which detection of the deoxyribonucleotides produced by RR occurs via coupling to the DNA polymerase reaction, and is enhanced by using RNase to degrade endogenous RNA. Cell extracts from various cell lines were treated with RNase and then reacted with ATP and radioactive ribonucleotide diphosphate as the substrate. Incorporation of the radioactive substrate [14C]CDP into DNA was linear over 30 min and was linear with the amount of extract, which provided RR activity. The...
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影响因子:2.458 (2012)

  • hydroxyurea 羟基脲
  • ribonucleotide 核糖核苷酸
  • radioactive 放射性的
  • RNase 核糖核酸酶
  • extracts 摘记
  • superior 优秀的
  • reaction 反酌
  • diphosphate 双磷盐酸
  • activity 活度
  • regulatory 控制的