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Updated: Jun 15, 2026

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Assessing Cellular Target Engagement by SHP2 (PTPN11) Phosphatase Inhibitors
Published on: July 17, 2020
通过M.结核病蛋白氨酸酸酶PtpB盖子域的动态活性部位保护
E Megan Flynn1, Jeffrey A Hanson, Tom Alber
1Department of Molecular and Biology and QB3 Institute, University of California, Berkeley, California 94720, USA.
Journal of the American Chemical Society
|March 17, 2010
概括
结核菌的蛋白质氨酸酸酶 PtpB 通过动态盖子机制抵抗氧化应激. 这个盖子快速打开和关闭,保护活动部位并减缓失活.
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:
- 结核菌菌的蛋白氨酸酸酶 PtpB (PtpB) 在宿主巨细胞中表现出对氧化应激的抵抗力.
- 这种氧化抵抗背后的机制尚未完全理解.
- 之前的晶体结构表明,PtpB有一个覆盖其活性部位的双螺旋盖.
研究的目的:
- 研究PtpB盖的动态及其在氧化抵抗中的作用.
- 阐明PtpB适应氧化条件的分子机制.
主要方法:
- 单分子福斯特型共振能量转移 (smFRET) 用于研究构成PtpB盖的两个螺旋体的动态.
- 进行了对开盘和收盘率的测量.
- 评估了溶剂粘度 (甘油和PEG8000) 对氧化失活率的影响.
主要成果:
- 获得了PtpB盖的自发打开和关闭过渡的直接证据.
- 盖子的封闭形状被偏爱了大约3: 1.
- 两个盖子螺旋异步移动,具有不同的打开和关闭速度.
- Lid 动态量化解释了氧化失活的缓慢速度.
- 增加溶剂粘度与糖醇,但不是PEG8000,通过增加开放的适应者群体加速氧化失活.
结论:
- PtpB盖子表现出快速的形状关,作为可逆物理封锁.
- 这种动态的盖子机制暂时掩盖了活性部位,减缓了氧化失活.
- PtpB对氧化应激的适应涉及其活性部位的保护性结构灵活性.
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