APE1活性部位残留物Asn174稳定了AP部位,对于催化是必不可少的
bioRxiv : the preprint server for biology
|January 27, 2025
概括
阿普里尼克/阿普里米迪尼克 (AP) 位点内核酶1 (APE1) 残留物N174对于DNA修复至关重要. 破坏其与AP位点相互作用的突变阻碍了催化,揭示了N1744.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 结构生物学 结构生物学
背景情况:
- 阿普里尼克/阿普里米尼克 (AP) 位点是常见的,突变性DNA病变.
- 基地切割修复 (BER) 路径地址为AP站点.
- 阿普里尼克/阿普里米尼克内核酶1 (APE1) 通过内核酶活性启动AP位点修复.
研究的目的:
- 研究APE1活性部位残留物N174.4的功能作用.
- 确定N174与AP位点的相互作用如何影响催化.
- 阐明APE1在处理AP站点中的分子机制.
主要方法:
- 局部定向的突变发生产生N174A,N174D和N174QAPE1突变.
- 酶分析测量野生类型和突变APE1.1的催化活性.
- 用X射线晶体学和计算模拟来分析蛋白质-DNA相互作用.
主要成果:
- APE1 N174A和N174D突变体因结合和静电相互作用的破坏导致催化功能受损.
- APE1 N174Q突变体表现出较少受损的催化作用,保留了关键相互作用.
- 结构和计算数据表明N174A和N174D突变体中的AP位点不稳定.
结论:
- 残留物N174对于稳定APE1活性位点内的AP位点至关重要.
- 涉及N174的结合和静电相互作用对于APE1催化非常重要.
- 这些发现为APE1在DNA修复中的作用提供了分子洞察力.
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