在ABC蛋白质中估计了前水解向外状态的真实稳定性
Márton A Simon1,2,3, Iordan Iordanov1,2,3, Andras Szollosi1,2,3
1Department of Biochemistry, Semmelweis University, Budapest, Hungary.
eLife
|October 2, 2023
概括
确定了囊性纤维化转膜导电调节器 (CFTR) 离子通道的非水解性闭合率. 破坏ATP水解的突变揭示了CFTR中核酸结合域界面的独特特征.
科学领域:
- 生物化学 生化学
- 分子生物学分子生物学
- 离子通道生理学 离子通道生理学
背景情况:
- 囊性纤维化跨膜导电调节器 (CFTR) 是一个ABC蛋白和离子通道.
- CFTR的ATP驱动的构造周期和孔隙开放爆发与核酸结合域 (NBD) 分解有关.
- ATP水解通常会终止CFTR开放爆发,但非水解性关闭速率仍然不太清楚.
研究的目的:
- 为了确定野生类型 (WT) CFTR的真正非水解性关闭率.
- 调查特定突变在破坏ATP水解和影响CFTR关中的作用.
- 发现人类CFTR中NBD接口的独特特征.
主要方法:
- 利用补丁记录来观察CFTR的单分子行为.
- 引入的特定突变 (K1250A,D1370N,E1371S,E1371Q) 破坏了ATP的水解.
- 测量和比较不同突变的突破持续时间和非水解性关闭率.
主要成果:
- K1250A,D1370N,E1371S和E1371Q突变完全取消了ATP的水解.
- 发现WT CFTR的真正非水解性关闭率与K1250A和E1371S突变的关闭率相似.
- 由于NBD间的H键,E1371Q突变使非水解率减慢了约15倍,而D1370N则加速了约15倍.
结论:
- 确定了WT CFTR的非水解性关闭率.
- 证明特定突变对NBD接口和ATP水解有不同的影响.
- 突出了人类CFTR NBD接口的独特结构和功能方面.
关键词:
这是一个D-循环.复合ATP-结合部位是复合的ATP结合部位.闪 关闭 关闭分子生物物理学分子生物物理学突变周期的突变周期结构生物学结构生物学异国主义者 (Xenopus) 是斑马鱼是一种斑马鱼.更多相关视频
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