酸化状态调节的HSP70结合:结构见解和补偿蛋白质工程
Mariah Stewart1,2, Chathura Paththamperuma3, Colleen McCann1
1The McAllister Heart Institute, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
bioRxiv : the preprint server for biology
|March 3, 2025
概括
热冲击蛋白70 (HSP70) 在T636的酸化削弱了它与E3酶CHIP的相互作用,影响了蛋白质质量控制. 这一发现为与蛋白质稳定失衡相关的疾病提供了洞察力.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 蛋白质质量控制对于细胞平衡至关重要,它依赖于诸如热冲击反应,无素-蛋白酶体系统和自-溶酶体通路等系统.
- 热冲击蛋白70 (HSP70) 和热冲击相关蛋白70 (HSC70) 是管理蛋白质折叠和降解的关键伴侣.
- 合作伙伴CHIP作为E3结合酶,准蛋白质通过ubiquitination降解.
研究的目的:
- 研究HSP70的C端酸化对其与CHIP的相互作用的影响.
- 阐明HSP70酸化对伴侣介导蛋白质降解的结构和功能后果.
- 探索这些相互作用对细胞平衡和疾病发病的影响.
主要方法:
- 利用无细胞和基于细胞的测定方法,包括X射线晶体学和生物层干扰计.
- 采用活细胞生物补充试验来研究细胞环境中的蛋白质相互作用.
- 设计了一种CHIP变体 (CHIP-G132N),以评估其与化HSP70.0的结合亲和力.
主要成果:
- 化HSP70 T636显著降低了与CHIP的结合亲和力,有利于其他辅助剂,如HOP.
- 结构分析显示,酸化破坏了关键的键,改变了HSP70和CHIP之间的结合动态.
- 虽然CHIP-G132N部分恢复了与酸化HSP70的结合,但其有效性在全长结构中降低了,这表明了复杂的调节相互作用.
结论:
- 翻译后的修改,特别是酸化,在调节蛋白质质量控制网络中的Chaperone-Co-Chaperone相互作用方面发挥着至关重要的作用.
- 由于酸化而改变的HSP70-CHIP相互作用会影响蛋白质的稳定性和降解,可能与癌症和神经退行性疾病有关.
- 了解这些分子机制为开发与蛋白质稳定性破坏相关的疾病的新疗法提供了基础.
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