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寨卡病毒NS2B-NS3蛋白酶的双重功能

Sergey A Shiryaev¹, Piotr Cieplak¹, Anton Cheltsov²

¹Sanford-Burnham-Prebys Medical Discovery Institute, Infectious and Inflammatory Disease Center, La Jolla, California, United States of America.

|November 27, 2023

概括

寨卡病毒NS2B-NS3蛋白酶将RNA结合在一个开放的形状,而不是封闭或超开放的状态. 这表明病毒酶-蛋白酶复合体具有新的RNA结合作用.

科学领域:

病毒学病毒学
分子生物学分子生物学
结构生物学结构生物学

背景情况:

寨卡病毒 (ZIKV) 血清蛋白酶对病毒复制至关重要,包括NS2B和NS3pro.
NS3酶域 (NS3hel) 对于酶活性和ATP结合至关重要.

研究的目的:

为了研究ZIKV NS2B-NS3pro.的新型RNA结合功能.
阐明参与RNA结合和蛋白酶活性的NS2B-NS3pro的构造状态.

主要方法:

生物化学试验测试ZIKV NS2B-NS3pro.pro的RNA结合亲和力 (Kd).
不同形状的NS2B-NS3pro的结构分析 (封闭,开放,超开放).
使用同类登革热病毒NS2B-NS3pro.的开放形状的建模.

主要成果:

ZIKV NS2B-NS3pro结合单链RNA,其Kd为0.3μM左右.
在封闭 (蛋白质活性) 和超开放的形状中,RNA结合被废除.
RNA结合发生在开放的构造中,由保存的正电荷结构促进.

结论:

ZIKV NS2B-NS3pro具有一种新的RNA结合功能,这取决于它的开放形状.
为交织的酶-蛋白酶机制提出了一个"反向虫"模型,涉及构造循环.
由基质诱导的过渡到闭合形状使蛋白酶活动成为可能.

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