Characterization of molecular interactions between Zika virus protease and peptides derived from the C-terminus of NS2B
Characterization of molecular interactions between Zika virus protease and peptides derived from the C-terminus of NS2B
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DOI：
10.1016/j.bbrc.2018.06.062
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发表时间：
2018-09-05
影响因子：
3.1
通讯作者：
Kang, CongBao
中科院分区：
文献类型：
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作者：
Li, Yan;Loh, Ying Ru;Kang, CongBao
寨卡病毒(Zika Virus，ZIKV)蛋白水解酶是一个双组分复合体，其中NS3含有催化三联体，NS2B辅助因子区域对酶的折叠和活性起重要作用。设计了一种不含NS2B跨膜区的蛋白水解酶载体eZiPro。对eZiPro的结构研究表明，NS2B C末端的Thr-GlyLys-Arg(TGKR)序列在切割后与活性部位结合。构建的bZiPro只含有NS2B辅因子区和NS3的N-末端，没有任何人工连接子或蛋白酶裂解位点，导致底物和抑制剂结合的空洞。在这里，我们证明了eZiPro中NS2B的TGKR序列是动态的。不同长度的NS2B多肽与bZiPro的结合亲和力不同。TGKR与eZiPro中活性部位的结合不影响蛋白酶与小分子化合物的结合。我们的结果表明，eZiPro也可用于评估小分子蛋白酶抑制剂。(C)2018 Elsevier Inc.保留所有权利。
Zika virus (ZIKV) protease is a two-component complex in which NS3 contains the catalytic triad and NS2B cofactor region is important for protease folding and activity. A protease construct-eZiPro without the transmembrane domains of NS2B was designed. Structural study on eZiPro reveals that the Thr-GlyLys-Arg (TGKR) sequence at the C-terminus of NS2B binds to the active site after cleavage. The bZiPro construct only contains NS2B cofactor region and the N-terminus of NS3 without any artificial linker or protease cleavage site, giving rise to an empty pocket accessible to substrate and inhibitor binding. Herein, we demonstrate that the TGKR sequence of NS2B in eZiPro is dynamic. Peptides from NS2B with various lengths exhibit different binding affinities to bZiPro. TGKR binding to the active site in eZiPro does not affect protease binding to small-molecule compounds. Our results suggest that eZiPro will also be useful for evaluating small-molecule protease inhibitors. (C) 2018 Elsevier Inc. All rights reserved.