Blocking human enterovirus 71 replication by targeting viral 2A protease
Blocking human enterovirus 71 replication by targeting viral 2A protease
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DOI：
10.1093/jac/dks304
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发表时间：
2012-12-01
影响因子：
5.2
通讯作者：
Violot, Sebastien
中科院分区：
文献类型：
--
作者：
Falah, Nisrine;Montserret, Roland;Violot, Sebastien
人类肠道病毒71型（EV-71）是肠道病毒属的一员，在亚太地区构成一个重大公共卫生问题，与若干严重的神经系统并发症有关。目前还没有针对EV-71的有效疫苗或抗病毒药物。这项研究的目的是确定6个氨基酸肽LVLQTM是否可以更广泛地用于肠道病毒，特别是EV-71。LVLQTM先前被证明可以抑制人鼻病毒（HRV） 2A蛋白酶（2A(pro)）的体外活性和HRV在小鼠体内的复制。为了研究LVLQTM肽是否是EV-71 2A（pro）的假底物，设计了含有LVLQTM序列的重组荧光素酶，以便2A（pro）识别该序列导致荧光素酶活化。等温滴定量热法进一步证实了EV-71 2A（pro）与LVLQTM肽的直接相互作用。然后，我们测试了肽对EV-71 2A（pro）切割活性和EV-71在HeLa细胞中的复制的影响。我们发现LVLQTM肽是EV-71 2A（pro）的有效底物类似物，它以9.6 m的解离速率结合到蛋白酶的活性位点，而且LVLQTM显著抑制2A（pro）的eIF4G裂解活性以及EV-71在HeLa细胞中的复制。本研究表明，先前被证明可以抑制HRV复制的LVLQTM肽也是EV-71 2A（pro）的有效抑制剂，因此可以抑制EV-71的复制，为开发新的EV-71抗病毒药物打开了新的大门。
Human enterovirus 71 (EV-71), a member of the Enterovirus genus, constitutes a major public health issue in the Asia-Pacific region, where it is associated with several severe neurological complications. There is currently no effective vaccine or antiviral against EV-71. The aim of this study was to determine whether the six amino acid peptide LVLQTM, which was previously shown to inhibit human rhinovirus (HRV) 2A protease (2A(pro)) activity in vitro and HRV replication in vivo in mice, could be of more general use against enteroviruses and more particularly against EV-71.To investigate whether the LVLQTM peptide was a pseudosubstrate of EV-71 2A(pro), a recombinant luciferase containing the LVLQTM sequence was designed so that recognition of this sequence by 2A(pro) led to luciferase activation. Direct interaction between EV-71 2A(pro) and the LVLQTM peptide was further confirmed by isothermal titration calorimetry. We then tested the effects of the peptide on EV-71 2A(pro) cleavage activity and EV-71 replication in HeLa cells.We showed that the LVLQTM peptide behaved as an effective substrate analogue of EV-71 2A(pro), which binds into the active site of the protease with a dissociation rate constant of 9.6 M. Moreover, LVLQTM significantly inhibited eIF4G cleavage activity of 2A(pro) as well as EV-71 replication in HeLa cells.This study demonstrates that the LVLQTM peptide that has previously been shown to inhibit HRV replication is also an effective inhibitor of EV-71 2A(pro) and therefore of EV-71 replication, opening new doors in the development of new antivirals against EV-71.