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Analysis of Nucleoside Triphosphate Hydrolysis by Middle East Respiratory Syndrome Coronavirus Helicase
Bulletin of the Korean Chemical Society ( IF 1.7 ) Pub Date : 2021-01-22 , DOI: 10.1002/bkcs.12229
Hye‐Min Woo 1, 2 , So‐Hee Pyo 1 , Yong‐Joo Jeong 1
Affiliation  

The purpose of this study is to measure the nucleoside triphosphate (NTP) hydrolysis by Middle East Respiratory Syndrome coronavirus (MERS‐CoV) helicase in the presence of different types of nucleic acids (NAs) for the determination of the optimal condition of hydrolysis activity. The steady‐state NTP hydrolysis analysis demonstrated that the MERS‐CoV helicase only (and considerably) hydrolyzed the ATP in the presence of poly(U). On the contrary, only UTP was considerably hydrolyzed in the presence of poly(C). When we used the DNAs as a substrate, the single‐stranded M13 DNA considerably stimulated the ATP and UTP hydrolysis. However, the ATP hydrolysis was not detected in the presence of ds DNA. The pH‐dependent ATP hydrolysis analysis demonstrated that the helicase optimally hydrolyzed ATP at a pH value of 7.0. The ATP hydrolysis was also performed in the presence of four different kinds of divalent cations, which demonstrated that only Mg2+ ions stimulated the ATP hydrolysis activity.

中文翻译:

中东呼吸综合征冠状病毒解旋酶水解三磷酸核苷的分析

这项研究的目的是在存在不同类型的核酸(NAs)的情况下,通过中东呼吸综合征冠状病毒(MERS-CoV)解旋酶测量三磷酸核苷(NTP)的水解,以确定最佳的水解活性条件。稳态NTP水解分析表明,在存在poly(U)的情况下,MERS-CoV解旋酶仅(且相当)水解了ATP。相反,在聚(C)的存在下,只有UTP被大量水解。当我们使用DNA作为底物时,单链M13 DNA极大地刺激了ATP和UTP水解。但是,在ds DNA存在下未检测到ATP水解。pH依赖的ATP水解分析表明,解旋酶在7.0的pH值下可以最佳地水解ATP。2+离子刺激了ATP的水解活性。
更新日期:2021-01-22
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