Profiling of substrate specificities of 3C-like proteases from group 1, 2a, 2b, and 3 coronaviruses.
Profiling of substrate specificities of 3C-like proteases from group 1, 2a, 2b, and 3 coronaviruses.
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DOI：
10.1371/journal.pone.0027228
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Wong KB
中科院分区：
文献类型：
--
作者：
Chuck CP;Chow HF;Wan DC;Wong KB
冠状病毒(CoV)根据其蛋白序列的多样性可分为甲型冠状病毒(第1组)、乙型冠状病毒(第2组)和伽马冠状病毒(第3组)。它们的3C型蛋白水解酶(3CLpro)催化多蛋白的蛋白分解过程以进行病毒复制，是一个潜在的抗冠状病毒感染的靶点。在这里，我们通过测量人冠状病毒NL63(组1)、人冠状病毒OC43(组2a)、严重急性呼吸综合征冠状病毒(SARS-CoV)(组2b)和传染性支气管炎病毒(IBV)(组3)的3CLpro对19×8个变异体的底物文库的活性来描述它们的底物特异性。结果与侧链体积、疏水性和取代残基的二级结构倾向等结构性质相关。所有3CLPro都偏好P1位的Gln，P2位的Leu，P3位的碱性残基，P4位的小疏水残基，以及P1‘和P2’位的小残基。尽管来自不同冠状病毒组的3CLPro在底物特异性上有许多相似之处，但在P4位观察到底物特异性的差异，IBV 3CLPro偏好P4-Pro，SARS-CoV 3CLPro偏好P4-Val。通过结合P3到P5位最有利的残基，我们确定了具有超活性的底物序列‘VARLQ↓sgf’和‘VPRLQ↓sgf’，其中VARLQ IBVsgf可以被所有3CLPro有效地切割，其相对活性为1.7%到3.2.本研究描述了来自第1、2a、2b和3Cov组的3CLpro的全面底物特异性，这可能为合理设计以蛋白酶为靶标的广谱模拟肽类抑制剂提供见解。
Coronaviruses (CoVs) can be classified into alphacoronavirus (group 1), betacoronavirus (group 2), and gammacoronavirus (group 3) based on diversity of the protein sequences. Their 3C-like protease (3CLpro), which catalyzes the proteolytic processing of the polyproteins for viral replication, is a potential target for anti-coronaviral infection. Here, we profiled the substrate specificities of 3CLpro from human CoV NL63 (group 1), human CoV OC43 (group 2a), severe acute respiratory syndrome coronavirus (SARS-CoV) (group 2b) and infectious bronchitis virus (IBV) (group 3), by measuring their activity against a substrate library of 19×8 of variants with single substitutions at P5 to P3' positions. The results were correlated with structural properties like side chain volume, hydrophobicity, and secondary structure propensities of substituting residues. All 3CLpro prefer Gln at P1 position, Leu at P2 position, basic residues at P3 position, small hydrophobic residues at P4 position, and small residues at P1' and P2' positions. Despite 3CLpro from different groups of CoVs share many similarities in substrate specificities, differences in substrate specificities were observed at P4 positions, with IBV 3CLpro prefers P4-Pro and SARS-CoV 3CLpro prefers P4-Val. By combining the most favorable residues at P3 to P5 positions, we identified super-active substrate sequences ‘VARLQ↓SGF’ that can be cleaved efficiently by all 3CLpro with relative activity of 1.7 to 3.2, and ‘VPRLQ↓SGF’ that can be cleaved specifically by IBV 3CLpro with relative activity of 4.3. The comprehensive substrate specificities of 3CLpro from each of the group 1, 2a, 2b, and 3 CoVs have been profiled in this study, which may provide insights into a rational design of broad-spectrum peptidomimetic inhibitors targeting the proteases.
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DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
DOI：
10.1016/s0140-6736(03)13967-0
发表时间：
2003-07-26
期刊：
Lancet (London, England)
影响因子：
--
作者：
Kuiken T;Fouchier RA;Schutten M;Rimmelzwaan GF;van Amerongen G;van Riel D;Laman JD;de Jong T;van Doornum G;Lim W;Ling AE;Chan PK;Tam JS;Zambon MC;Gopal R;Drosten C;van der Werf S;Escriou N;Manuguerra JC;Stöhr K;Peiris JS;Osterhaus AD
通讯作者：
Osterhaus AD
影响因子：
3.8
作者：
Hegyi, A;Ziebuhr, J
通讯作者：
Ziebuhr, J
影响因子：
56.9
作者：
Anand, K;Ziebuhr, J;Hilgenfeld, R
通讯作者：
Hilgenfeld, R
影响因子：
7.6
作者：
Li SY;Chen C;Zhang HQ;Guo HY;Wang H;Wang L;Zhang X;Hua SN;Yu J;Xiao PG;Li RS;Tan X
通讯作者：
Tan X