Vital NS3 helicase activity is inhibited by peptides reproducing the Arg-rich conserved motif of the enzyme (motif VI)
Vital NS3 helicase activity is inhibited by peptides reproducing the Arg-rich conserved motif of the enzyme (motif VI)
复制标题
DOI：
10.1016/j.bcp.2008.03.018
复制
发表时间：
2008-07-01
影响因子：
5.8
通讯作者：
Baier, Andrea
中科院分区：
文献类型：
--
作者：
Borowski, Peter;Heising, Maite V.;Baier, Andrea
流感病毒的NTPase/解旋酶是其复制复合体的重要组成部分。该酶由7个高度保守的氨基酸基序组成。随机筛选大量丙型肝炎病毒(丙型肝炎病毒)衍生的多肽，发现与丙型肝炎病毒NTPase/解旋酶的基序VI相对应的碱性氨基酸延伸(丙型肝炎病毒多蛋白的1487-1500氨基酸)。合成了与丙型肝炎病毒、西尼罗河病毒(WNV)和日本脑炎病毒(JEV)基序相对应的多肽，作为NTPase和病毒酶介导的解离反应的抑制剂。在长度和结构上有区别的多肽。在所测试的多肽中，复制基序VI序列的丙型肝炎病毒(14871500)是对所研究酶的解旋酶活性最有效的抑制物。其他相应的多肽是相当温和的抑制剂。所研究的多肽对流感病毒科解旋酶的抑制作用依次为：丙型肝炎病毒(1487-1500)和西尼罗河病毒(1959-1572)和乙脑病毒(1962-1975)。有趣的是，解旋酶活性对多肽抑制的敏感性是相似的，顺序为：丙型肝炎病毒>；西尼罗河病毒>；JEV。抑制是由于NTP结合和水解位以外的酶活性部位的结合和阻断所致。动力学分析表明，这些多肽的结合不影响酶的NTPase活性。该多肽可以作为有效的、选择性的工具来抑制病毒的繁殖。(C)2008 Elsevier Inc.保留所有权利。
The NTPase/helicase of Flauiviridae viruses is one of the essential components of their replication complex. The enzyme is defined by the presence of seven highly conserved amino acid motifs. Random screening of numerous hepatitis C virus (HCV) derived peptides, revealed a basic amino acid stretch corresponding to motif VI of the HCV NTPase/helicase (amino acids 1487-1500 of the HCV polyprotein). This peptide inhibited the unwinding activity of the enzyme with an IC50 = 0.2 mu M. Peptides corresponding to motif VI of HCV, West Nile virus (WNV) and Japanese encephalitis virus (JEV) were synthesized and tested as inhibitors of NTPase and unwinding reactions mediated by the viral enzymes. Peptides distinguished in regard to their length and structure. Between the peptides tested HCV(14871500) reproducing the sequence of motif VI was the most potent inhibitor of helicase activities of investigated enzymes. Other respective peptides were rather modest inhibitors. The examined peptides inhibited the Flauiviridae helicases in the following order of potency: HCV(1487-1500) > WNV(1959-1572) > JEV(1962-1975). Interestingly, the susceptibility of the helicase activity to the inhibition by the peptides was similar and in the row: HCV > WNV > JEV.The inhibition results from binding and blockade of the active site of the enzyme lyes beyond the NTP-binding and hydrolyzing site. The kinetic analyses indicated that the binding of the peptides do not interfere with the NTPase activity of the enzymes. The peptide may serve as effective and selective tool to reduce the virus propagation. (c) 2008 Elsevier Inc. All rights reserved.