通过甲基-TROSY NMR研究的卡斯帕-9在光体上的激活
Alexander I M Sever1,2, T Reid Alderson1,3,4, Enrico Rennella1,3,4
1Department of Chemistry, University of Toronto, Toronto, ON M5S 3H6, Canada.
概括
光体支架组织caspase-9蛋白酶域,保持它们单质,直到基质结合触发二元化和激活. 这揭示了编程细胞死亡的关键调节步骤,对于理解癌症逃避细胞亡至关重要.
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:
- 线粒体的亡信号传递涉及到亡的亡,激活caspase-9.
- 卡斯巴-9激活对于编程细胞死亡至关重要,但在癌症中经常被规避.
- 卡斯巴-9在细胞核上激活的结构机制尚不清楚.
研究的目的:
- 阐明在细胞上酶-9激活的结构机制.
- 为了研究细胞体支架在调节caspase-9活动中的作用.
主要方法:
- 用甲基横向放松优化的NMR光谱学研究了caspase-9.
- 用蛋白质工程和生物化学测试来分析卡斯帕-9-阿波托索姆相互作用.
- 核磁共振被应用于原生1.3-MDa光体复合体和工程仿真体.
主要成果:
- 卡斯巴酶-9及其蛋白酶域 (PD) 单独表现出弱二分化.
- 卡斯帕-9 PD 在被招募到阿波托索姆支架时仍然是单质的.
- 细胞质体结合只在基质存在时,便于快速和广泛的caspase-9 PD二分化.
结论:
- Апопто组织卡斯帕-9 酶,使基质诱导的二分化和激活成为可能.
- 这提供了一个关键的调节机制来控制被编程的细胞死亡.
- 核磁共振光谱对于特征大型多MDa蛋白质复合体,如光体,是有效的.
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