Conservation of substrate specificities among coronavirus main proteases
Conservation of substrate specificities among coronavirus main proteases
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DOI：
10.1099/0022-1317-83-3-595
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发表时间：
2002-03-01
影响因子：
3.8
通讯作者：
Ziebuhr, J
中科院分区：
文献类型：
--
作者：
Hegyi, A;Ziebuhr, J
冠状病毒复制酶多蛋白加工中的关键酶是冠状病毒主酶3CL(Pro)。对包括冠状病毒I、II和III组的原型酶在内的5种冠状病毒主要蛋白酶的底物特异性进行了研究。对人冠状病毒(HCoV)、传染性胃肠炎病毒(TGEV)、猫传染性腹膜炎病毒、禽传染性支气管炎病毒和小鼠肝炎病毒(MHV)的重组主要蛋白酶进行了基于多肽的反式切割分析。一组相应的HCoV、TGEV和MHV 3CL(Pro)裂解位点的相对速率常数的测定揭示了这些位点的保守排名。此外，一个代表HCoV3CL(Pro)N端裂解位点的合成肽被非同源的主要蛋白水解酶有效地降解。这些数据表明，pp1a/pp1ab内位点的差异切割动力学是冠状病毒主要蛋白酶的保守特征，并使我们预测所有冠状病毒复制酶多蛋白的加工动力学相似。
The key enzyme in coronavirus replicase polyprotein processing is the coronavirus main protease, 3CL(pro). The substrate specificities of five coronavirus main proteases, including the prototypic enzymes from the coronavirus groups I, II and III, were characterized. Recombinant main proteases of human coronavirus (HCoV), transmissible gastroenteritis virus (TGEV), feline infectious peritonitis virus, avian infectious bronchitis virus and mouse hepatitis virus (MHV) were tested in peptide-based trans-cleavage assays. The determination of relative rate constants for a set of corresponding HCoV, TGEV and MHV 3CL(pro) cleavage sites revealed a conserved ranking of these sites. Furthermore, a synthetic peptide representing the N-terminal HCoV 3CL(pro) cleavage site was shown to be effectively hydrolysed by non-cognate main proteases. The data show that the differential cleavage kinetics of sites within pp1a/ pp 1 ab are a conserved feature of coronavirus main proteases and lead us to predict similar processing kinetics for the replicase polyproteins of all corona-viruses.