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施莱芬11 N-终端域的结构和生化表征

Pengjiao Hou^1,2, Wei Hao^1,2, Bo Qin^1,2

¹NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, PR China.

Nucleic acids research

|June 9, 2023

概括

睡眠11 (SLFN11) 蛋白质是一种

科学领域:

结构生物学是结构生物学.
分子生物学分子生物学
生物化学生化学

背景情况:

施莱芬11 (SLFN11) 是癌症治疗和病毒与宿主相互作用中的关键蛋白质.
了解SLFN11的结构和功能对于治疗开发至关重要.

研究的目的:

确定Sus scrofa SLFN11 N端域 (NTD) 的晶体结构.
研究sSLFN11-NTD的生物化学特征和酶活性.
阐明SLFN11在翻译抑制中的作用的结构基础.

主要方法:

进行X射线晶体学以确定sSLFN11-NTD的2.69 Å分辨率结构.
在体外RNase测试以评估各种RNA类型 (tRNA,rRNA) 的分裂.
位点定向突变发生,以确定影响核分解活性的关键残留物和结构元素.
基于细胞的测试来评估SLFN11对蛋白质翻译的影响.

主要成果:

sSLFN11-NTD 的晶体结构揭示了具有明显生化特性的形折叠.
sSLFN11-NTD作为一种强大的RNase功能,优先分裂II型tRNA和rRNA.
突变分析确定了连接循环和特定残留物 (例如,E42) 对于RNase活动和基质识别至关重要.
E42A突变增强了RNase活性,而E209A突变取消了翻译抑制,突出了NTD的作用.

结论:

对sSLFN11-NTD的结构和生化特征提供了对施莱芬家族机制的洞察.
SLFN11的RNase活动,特别是NTD的RNase活动,对于其在基于编码子使用的翻译抑制中的作用至关重要.
像E42和E209这样的关键残留物是SLFN11酶功能和细胞效应的关键决定因素.

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