实时动态跟踪活细胞中多个基除修复酶的实时动态跟踪
Junqiu Zhai1, Han Zhang1, Wenzhi Zhu1
1School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
Analytical chemistry
|February 27, 2025
概括
科学家们开发了一种新的DNA探针,用于跟踪活细胞中的基切除修复 (BER) 酶,apurinic/apyrimidinic内核酶1 (APE1) 和内核酶1 (FEN1). 这个工具可以检测环境污染物的DNA损伤,如1-MP和6-Cl-BaP.
科学领域:
- 生物化学 生化学
- 分子生物学分子生物学
- 环境科学 环境科学
背景情况:
- 同时监测基切除修复 (BER) 酶,apurinic/apyrimidinic内核酶1 (APE1) 和内核酶1 (FEN1),对于了解疾病发展和污染物诱导的细胞毒性至关重要.
- 目前的方法在全面分析这些酶在活细胞中的多样性功能方面是有限的.
研究的目的:
- 开发一种可双激活的新型DNA光探针 (AP-FLAP),用于同时在现场可视化APE1和FEN1活动.
- 揭示和量化生活细胞中环境污染物引起的与BER相关的DNA损伤.
主要方法:
- 设计了一种可双激活的DNA探头 (AP-FLAP),集成子和发针结构,在酶裂变时释放不同的光信号 (APE1的ROX,FEN1的FAM).
- 将AP-FLAP探针应用于暴露于1-甲基 (1-MP) 和6-enzo[a]pyrene (6-Cl-BaP) 的人静脉内皮细胞 (HUVEC).
主要成果:
- AP-FLAP探测器展示了APE1和FEN1活动的特定和敏感检测.
- 在暴露于1-MP和6-Cl-BaP的HUVEC中观察到显著的基损伤,与污染物度相关.
- 即使在低度 (0.1μM) 中,6-Cl-BaP也会以剂量依赖的方式诱导显著的DNA损伤.
结论:
- AP-FLAP 探针是一种强大的工具,用于阐明BER分子机制和DNA损伤修复,以应对环境暴露.
- 这种方法为开发用于化学生物学和生物医学研究的多功能核酸探针开辟了新的途径.
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