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Sinduja K Marx¹, Keith J Mickolajczyk^2,3, Jonathan M Craig¹

¹Department of Physics, University of Washington, Seattle, WA 98195, USA.

Nucleic acids research

|August 10, 2023

概括

SARS-CoV-2 螺旋酶 (nsp13) 使用 ATP 水解来沿核酸运动,单分子纳米孔针揭示了其精确的步骤. 抑制研究表明ATPase抑制剂具有多种机制,取决于施加的力.

科学领域:

生物物理学的生物物理.
分子生物学分子生物学
病毒学病毒学

背景情况:

在SARS-CoV-2基因组编码的螺旋酶nsp13,至关重要的病毒复制和潜在的抗病毒目标.
了解NSP13的机制是开发有效抗病毒疗法的关键.

研究的目的:

通过纳米孔子来阐明SARS-CoV-2螺旋酶 (nsp13) 转位和解的单分子机制.
研究ATPγS对nsp13活性的抑制机制.

主要方法:

使用高分辨率单分子纳米孔子来测量nsp13活动.
分析了单链DNA上的nsp13转位和双链DNA的解.
在ATPase抑制剂ATPγS的存在下对nsp13运动进行了动态分析.

主要成果:

确定了nsp13的单核酸转位速度 (~1000 nt/s) 和解速度 (~100 bp/s).
揭示了ATPγS的多个抑制机制,依赖力.
建立了NSP13抑制的详细动力学模型.

结论:

纳米孔 tweezers 提供高分辨率的洞察力病毒酶的功能和抑制.
ATPγS表现出复杂的,取决于力量的nsp13抑制.
这项工作为病毒化酶抑制剂的单分子研究提供了基础.

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