化学诱导的多功能阿普里尼克/阿普里米尼克内核酶1的部分展开
bioRxiv : the preprint server for biology
|July 10, 2023
概括
阿普里尼克/阿普里米尼克内核酶I (APE1) 在受抑制剂APX3330的影响下,对其氧化还原功能进行可逆的部分展开. 这种展开暴露了关键的残留物,影响了转录因子调节.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- 阿普里尼克/阿普里米尼克内核酶I (APE1) 对于细胞生存至关重要,具有内核酶和氧化还原因子活动.
- 这些不同的功能需要APE1的不同构造状态:完全折叠用于内核酶活性,部分展开用于氧化还原活性.
- 反氧化活性涉及暴露Cys65以减少像AP-1,NF-κB和HIF-1α这样的转录因子,从而促进DNA结合.
研究的目的:
- 阐明APE1与其氧化还原功能相关的部分展开背后的分子机制.
- 描述氧化还原抑制剂APX3330与APE1的相互作用及其对APE1构造的影响.
- 了解由APX3330.30引起的APE1形状变化的可逆性.
主要方法:
- 使用WaterLOGSY和1H-15N HSQC NMR光谱来研究APE1与APX3330的相互作用.
- 实验中使用乙醇和乙二作为辅溶剂进行了实验,以探测形状变化.
- 化学转移干扰 (CSP) 分析和化温度测定被用于评估结合和稳定性.
主要成果:
- APX3330与APE1的内核酶活性部位和在乙基中的独特口袋结合,诱导部分展开 (1H-15N HSQC信号的~35%的损失).
- 部分展开的区域涉及N-和C-终端β链,表明了特定的结构重排.
- 结合APX3330可逆地降低APE1的化温度,而不会影响其内核酶活性.
结论:
- 这项研究为APE1的可逆部分展开提供了分子洞察力,这对其氧化还原功能至关重要.
- 这些发现澄清了APX3330的氧化还原抑制机制,突出了它与特定APE1构造的相互作用.
- 这项研究有助于了解APE1的双重活动是如何在结构层面上受到监管的.
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