在SARS-CoV-2RNA依赖RNA聚合酶中自然发生突变的生物化学特征
Matěj Danda1, Anna Klimešová1, Klára Kušková1
1Department of Biotechnology, University of Chemistry and Technology, Prague, Czech Republic.
Protein science : a publication of the Protein Society
|August 15, 2024
概括
在SARS-CoV-2RNA依赖RNA聚合酶 (RdRp) 子单元中的突变会影响其稳定性和功能. 一些突变增加了病毒聚合酶活性,为抗病毒药物开发提供了见解.
科学领域:
- 病毒学 病毒学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 在SARS-CoV-2RNA依赖RNA聚合酶 (RdRp) 中的突变很常见.
- RdRp是抗病毒疗法的关键目标.
- 关于RdRp亚单元突变的表型影响的实验数据有限.
研究的目的:
- 研究SARS-CoV-2 RdRp子单元 (nsp7,nsp8,nsp12) 中突变的表型影响.
- 评估这些突变对蛋白质稳定性,复杂组合和聚合酶活性的影响.
主要方法:
- 纳米差异扫描度测量用于测量蛋白质稳定性.
- 微尺度热泳以分析复杂的组件.
- 度酶定量测试以确定RNA聚合酶活性.
主要成果:
- 突变对蛋白质稳定性和nsp12结合亲和力产生了不同的影响.
- 一个nsp8突变增加了13°C的蛋白质稳定性.
- 大多数突变改变了聚合酶活性;双重突变 (nsp8 A21V/nsp12 P323L) 增加了50%的活性.
结论:
- 这项研究提供了所有SARS-CoV-2 RdRp子单元中突变的第一个生化分析.
- 观察到的稳定性和组合的变化与改变的聚合酶活性相关.
- 研究结果提供了对RdRp功能和SARS-CoV-2亚变体中潜在的治疗点的见解.
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