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SURFS和AlphaFold揭示了由EF-Tu D81突变引起的核糖体足迹转移

Yi Zeng, Jordan Johnson, Shoujun Xu

bioRxiv : the preprint server for biology

|June 12, 2025

概括

关键蛋白质延长因子-Tu (EF-Tu) 的突变破坏了它在蛋白质合成过程中释放转移RNA (tRNA) 的能力. 这会影响翻译效率和核糖体动态.

科学领域:

分子生物学分子生物学
生物化学生化学
结构生物学结构生物学

背景情况:

蛋白质合成的准确性取决于正确的信使RNA (mRNA) 通过转移RNA (tRNA) 进行解码.
延长因子-Tu (EF-Tu) 对于指导这种解码过程至关重要.
在EF-Tu中保存的残留物在其功能中起着至关重要的作用.

研究的目的:

为了研究突变对阿斯巴达酸81 (D81) 的功能影响,阿斯巴达酸81 (D81) 是EF-Tu.的保存残留物.
阐明D81突变影响EF-Tu功能和翻译保真性的分子机制.

主要方法:

测量GTPase活性测试以测量酶功能.
用于结构预测的AlphaFold建模 (AlphaFold2和AlphaFold3).
基于量子感应的超分辨率力谱学 (SURFS) 来探测核糖体动力学.
亚核酸核糖体足迹测定用于评估tRNA释放.

主要成果:

所有D81突变都保留了GTPase活性,但影响了从核糖体释放的tRNA.
AlphaFold3 建模表明,D81 突变破坏了与 GTP 结合状态下的离子协调和与素-素循环的相互作用.
AlphaFold2分析揭示了D81的共同进化约束,它的突变允许神秘的结构变化.

结论:

一个单一的保存残留物,D81,在EF-Tu中连接了催化协调,全osteric通信和进化约束.
在D81的突变提供了对翻译忠实性的机制性见解.
超分辨率力光谱 (SURFS) 是研究核糖体动态的一个有价值的工具.

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