Prediction and biochemical analysis of putative cleavage sites of the 3C-like protease of Middle East respiratory syndrome coronavirus.
Prediction and biochemical analysis of putative cleavage sites of the 3C-like protease of Middle East respiratory syndrome coronavirus.
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中东呼吸综合征冠状病毒 3C 样蛋白酶假定裂解位点的预测和生化分析。
DOI：
10.1016/j.virusres.2015.05.018
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发表时间：
2015-10-02
期刊：
影响因子：
5
通讯作者：
Guo D
中科院分区：
文献类型：
--
作者：
Wu A;Wang Y;Zeng C;Huang X;Xu S;Su C;Wang M;Chen Y;Guo D
提出了一种冠状病毒3CLpro切割位点的预测方法，以平衡准确性和假阳性。9个假定的非典型切割位点中的3个被验证，它们位于nsp 4的上游。确定了MERS-CoV 3CLpro的所有11个典型切割位点，并计算了米氏常数。冠状病毒3C样蛋白酶（3CLpro）负责切割冠状病毒多聚蛋白1a/1ab（pp 1a/1ab），以产生nsp 4 -16的成熟非结构蛋白（nsp）。新出现的中东呼吸综合征冠状病毒（MERS-CoV）的nsp 5被鉴定为3CLpro，并且基于序列比对预测了其典型切割位点（nsps之间），但是这些切割位点的可切割性仍有待实验证实，pp 1a/1ab内的推定非典型切割位点（在一个nsp内）有待进一步分析。在这里，我们提出了一种用于预测冠状病毒3CLpro切割位点的方法，该方法平衡了预测准确性和假阳性结果。通过将该方法应用于MERS-CoV，11个典型的切割位点很容易被鉴定并通过生化测定进行验证。MERS-CoV典型切割位点的米氏常数表明，MERS-CoV 3CLpro的底物特异性相对保守。有趣的是，预测了9个假定的非典型切割位点，其中3个可以被MERS-CoV nsp 5切割。这些结果为冠状病毒nsp新产物的鉴定和功能鉴定奠定了基础。
A prediction method of coronaviral 3CLpro cleavage sites was proposed to balance the accuracy and false positives. Three of the nine putative non-canonical cleavage sites were verified, which are located upstream to nsp4. All 11 canonical cleavage sites of MERS-CoV 3CLpro were confirmed and the Michaelis constants were calculated. Coronavirus 3C-like protease (3CLpro) is responsible for the cleavage of coronaviral polyprotein 1a/1ab (pp1a/1ab) to produce the mature non-structural proteins (nsps) of nsp4–16. The nsp5 of the newly emerging Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as 3CLpro and its canonical cleavage sites (between nsps) were predicted based on sequence alignment, but the cleavability of these cleavage sites remains to be experimentally confirmed and putative non-canonical cleavage sites (inside one nsp) within the pp1a/1ab awaits further analysis. Here, we proposed a method for predicting coronaviral 3CLpro cleavage sites which balances the prediction accuracy and false positive outcomes. By applying this method to MERS-CoV, the 11 canonical cleavage sites were readily identified and verified by the biochemical assays. The Michaelis constant of the canonical cleavage sites of MERS-CoV showed that the substrate specificity of MERS-CoV 3CLpro is relatively conserved. Interestingly, nine putative non-canonical cleavage sites were predicted and three of them could be cleaved by MERS-CoV nsp5. These results pave the way for identification and functional characterization of new nsp products of coronaviruses.
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影响因子：
14.9
作者：
Gorbalenya AE;Koonin EV;Donchenko AP;Blinov VM
通讯作者：
Blinov VM
中东呼吸综合征3C样蛋白酶的结构揭示了对底物特异性的见解。
DOI：
10.1107/s1399004715003521
发表时间：
2015-05
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Needle D;Lountos GT;Waugh DS
通讯作者：
Waugh DS
DOI：
10.1074/jbc.m310875200
发表时间：
2004-01-16
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Fan K;Wei P;Feng Q;Chen S;Huang C;Ma L;Lai B;Pei J;Liu Y;Chen J;Lai L
通讯作者：
Lai L
影响因子：
3.8
作者：
Hegyi, A;Ziebuhr, J
通讯作者：
Ziebuhr, J
影响因子：
5.4
作者：
Ziebuhr, John;Schelle, Barbara;Thiel, Volker
通讯作者：
Thiel, Volker