Critical Assessment of the Important Residues Involved in the Dimerization and Catalysis of MERS Coronavirus Main Protease.
Critical Assessment of the Important Residues Involved in the Dimerization and Catalysis of MERS Coronavirus Main Protease.
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DOI：
10.1371/journal.pone.0144865
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Chou CY
中科院分区：
文献类型：
--
作者：
Ho BL;Cheng SC;Shi L;Wang TY;Ho KI;Chou CY
沙特阿拉伯吉达等地出现一种高致病性人类冠状病毒（CoV）--中东呼吸综合征冠状病毒（MERS-CoV），自2012年9月起迅速向欧洲和亚洲国家蔓延。截至2015年10月1日，它已感染了至少1593人，全球死亡率约为35%。了解这种病毒的研究是必要和紧迫的。在本研究中，MERS-CoV的主要蛋白酶（Mpro）的表达，蛋白质的二聚体及其催化的关系进行了研究。MERS-CoV Mpro的晶体结构表明，它与其他冠状病毒Mpro具有相似的支架，由胰凝乳蛋白酶样结构域I和II以及五个螺旋的螺旋结构域III组成。分析性超离心分析表明，MERS-CoV Mpro在肽底物存在下经历单体至二聚体的转化。Glu 169是一个关键残基，在二聚和催化中起着双重作用。在二聚化界面上发现的其他残基的诱变表明MERS-CoV Mpro的二聚化是其催化活性所需的。一个突变，M298 R，导致在一个稳定的二聚体具有更高水平的蛋白水解活性比野生型酶。MERS-CoV Mpro显示底物诱导的二聚化和有效的蛋白水解活性。对这些过程重要的残基的关键评估提供了对冠状病毒Mpro家族内二聚化和催化之间相关性的见解。
A highly pathogenic human coronavirus (CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), has emerged in Jeddah and other places in Saudi Arabia, and has quickly spread to European and Asian countries since September 2012. Up to the 1st October 2015 it has infected at least 1593 people with a global fatality rate of about 35%. Studies to understand the virus are necessary and urgent. In the present study, MERS-CoV main protease (Mpro) is expressed; the dimerization of the protein and its relationship to catalysis are investigated. The crystal structure of MERS-CoV Mpro indicates that it shares a similar scaffold to that of other coronaviral Mpro and consists of chymotrypsin-like domains I and II and a helical domain III of five helices. Analytical ultracentrifugation analysis demonstrated that MERS-CoV Mpro undergoes a monomer to dimer conversion in the presence of a peptide substrate. Glu169 is a key residue and plays a dual role in both dimerization and catalysis. The mutagenesis of other residues found on the dimerization interface indicate that dimerization of MERS-CoV Mpro is required for its catalytic activity. One mutation, M298R, resulted in a stable dimer with a higher level of proteolytic activity than the wild-type enzyme. MERS-CoV Mpro shows substrate-induced dimerization and potent proteolytic activity. A critical assessment of the residues important to these processes provides insights into the correlation between dimerization and catalysis within the coronaviral Mpro family.