细胞质hPGK1的酸化影响蛋白质稳定性和连接体结合:对其在癌症中的亚细胞向的影响
Juan Luis Pacheco-García1, Mario Cano-Muñoz1, Dmitry S Loginov2
1Departamento de Química Física, Universidad de Granada, Spain.
The FEBS journal
|September 6, 2024
概括
在S256中对人类糖酸酶1 (hPGK1) 的酸化使酶不稳定,降低了Mg-ADP的结合亲和力. 这种蛋白质稳定性变化可能会增强hPGK1在癌症中的核转位.
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:
- 人类糖酸酶1 (hPGK1) 是一个关键的糖解酶,调节细胞ATP/ADP平衡.
- 在S203和S256的酸化影响hPGK1定位到线粒体和核,分别影响瘤发生.
- 亚细胞局部化变化与蛋白质稳定性的特定位点变化有关.
研究的目的:
- 研究特定位点酸化对hPGK1热和运动稳定性的影响.
- 使用计算和实验方法分析蛋白质结构动力学和连接物结合亲和性 (3-糖酸盐和Mg-ADP).
- 阐明hPGK1酸化诱导的稳定性变化的结构和能量基础.
主要方法:
- 使用了-交换 (HDX) 和分子动力学 (MD) 模拟.
- 进行热量测量研究以评估连接体结合热力学.
- 引入了相仿性突变 (S203D,S256D),以模仿酸化效应.
主要成果:
- S256D突变显著减少了hPGK1的动力稳定性50倍,而S203D没有影响.
- 对于S256D变种,观察到Mg-ADP结合亲和度的显著下降.
- HDX和MD模拟证实了S256D诱导的不稳定性和由于结合部位的不稳定性而降低了Mg-ADP亲和力.
结论:
- 特定位点的化,特别是S256,对hPGK1蛋白稳定性和Mg-ADP结合有深刻的影响.
- 这种S256D突变会破坏Mg-ADP结合部位的稳定,从而解释了这种降低的亲和力.
- 改变hPGK1稳定性可能会导致癌症中的核转位,尽管线粒体进口机制需要进一步研究.
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