一种来自大肠杆菌的DNA糖基化酶，可从含有碱基残基片段的多脱氧核糖核苷酸中释放游离尿素。
A DNA glycosylase from Escherichia coli that releases free urea from a polydeoxyribonucleotide containing fragments of base residues.
作者信息
Breimer L, Lindahl T
出版信息
Nucleic Acids Res. 1980 Dec 20;8(24):6199-211. doi: 10.1093/nar/8.24.6199.
Abstract
A poly (dA, [2-14C]dT) copolymer has been synthesized using terminal deoxynucleotidyltransferase. Treatment of the polydeoxyribonucleotide with potassium permanganate converts the thymine residues to urea and N-substituted urea derivatives, while the adenine residues are resistant to oxidation. This damaged polymer has been annealed with an equimolar amount of poly (dT) to generate a double-stranded polydeoxyribonucleotide containing scattered fragmented base residues, which are radioactively labeled selectively. On incubation of the latter with crude cell extracts from E. coli, free urea is released by a DNA glycosylase activity. The enzyme has been partly purified, and appears to be different from previously studied DNA glycosylase. It shows a strong preference for a double-stranded substrate, exhibits no cofactor requirement, and has a molecular weight of 20000 - 25000. Since fragmentation of pyrimidine residues is a major type of base lesion introduced in DNA by exposure to ionizing radiation, it seems likely this DNA glycosylase is active in repair of X-ray-induced lesions.
摘要
利用末端脱氧核苷酸转移酶合成了聚（dA，[2-¹⁴C]dT）共聚物。用高锰酸钾处理该聚脱氧核糖核苷酸会使胸腺嘧啶残基转化为尿素和N-取代尿素衍生物，而腺嘌呤残基对氧化具有抗性。这种受损的聚合物已与等摩尔量的聚（dT）退火，以生成含有分散的片段化碱基残基的双链聚脱氧核糖核苷酸，这些残基被选择性地放射性标记。将后者与大肠杆菌的粗细胞提取物一起温育时，一种DNA糖基化酶活性会释放出游离尿素。该酶已部分纯化，似乎与先前研究的DNA糖基化酶不同。它对双链底物有强烈偏好，不需要辅助因子，分子量为20000 - 25000。由于嘧啶残基的片段化是DNA暴露于电离辐射后引入的主要碱基损伤类型，这种DNA糖基化酶似乎在修复X射线诱导的损伤中具有活性。
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