大肠杆菌和人7,8-二氢-8-氧鸟嘌呤DNA糖基酶的酶学性质。

K Asagoshi, T Yamada, H Terato, Y Ohyama, H Ide
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引用次数: 6

摘要

DNA的氧化损伤会产生异常的鸟嘌呤碱基,如2,6-二氨基-4-羟基甲脒-嘧啶(Fapy)和7,8-二氢-8-氧鸟嘌呤(8-oxoG)。虽然含有单个8-oxoG的合成寡核苷酸已被广泛用于研究这种病变的酶促处理,但迄今为止还没有合成含有Fapy的寡核苷酸作为一种独特的病变。在本研究中,通过DNA聚合酶反应和随后的碱处理制备了含有单个2,6-二氨基-4-羟基-5-(n -甲基)甲脒的寡核苷酸(me-Fapy, Fapy的甲基化衍生物)。利用含有me-Fapy和8-oxoG的寡核苷酸底物比较了Fpg和hOGG1蛋白的修复活性。
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Enzymatic properties of Escherichia coli and human 7,8-dihydro-8-oxoguanine DNA glycosylases.

Oxidative damage to DNA generates aberrant guanine bases such as 2,6-diamino-4-hydroxy-formamido-pyrimidine (Fapy) and 7,8-dihydro-8-oxoguanine (8-oxoG). Although synthetic oligonucleotides containing a single 8-oxoG have been widely used to study enzymatic processing of this lesion, the synthesis of oligonucleotides containing Fapy as a unique lesion has not been achieved to date. In this study, an oligonucleotide containing a single 2,6-diamino-4-hydroxy-5-(N-methyl)formamido-pyrimidine (me-Fapy, a methylated derivative of Fapy) was prepared by a DNA polymerase reaction and the subsequent alkali treatment. The repair activity of Fpg and hOGG1 proteins were compared using oligonucleotide substrates containing me-Fapy and 8-oxoG.

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