Competitive inhibition of the dengue virus NS3 serine protease by synthetic peptides representing polyprotein cleavage sites
Competitive inhibition of the dengue virus NS3 serine protease by synthetic peptides representing polyprotein cleavage sites
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DOI：
10.1016/j.bbrc.2005.03.107
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发表时间：
2005-05-20
影响因子：
3.1
通讯作者：
Katzenmeier, G
中科院分区：
文献类型：
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作者：
Chanprapaph, S;Saparpakorn, P;Katzenmeier, G
登革热病毒的 NS3 丝氨酸蛋白酶是病毒多蛋白成熟所必需的，因此代表了抗病毒抑制剂开发的有希望的靶标。然而，该酶的底物特异性仅在有限程度上得到表征。在本研究中，我们研究了源自天然多蛋白底物的 P6-P1 和 P1'-P5' 区域的合成肽对 NS3 蛋白酶的产物抑制。发现对应于多蛋白 P6-P1 区域的 N 端裂解位点肽充当酶的竞争性抑制剂，其 K-i 值范围为 67 至 12 muM。发现代表 NS2A/NS2B 裂解位点的肽 RTSKKR 的 K-i 值最低。通过使用较短的肽、SKKR、KKR、KR、AGRR 和 GKR 分析该切割位点序列的抑制作用。除AGRR肽在1mM浓度下不抑制蛋白酶外，所有其他肽的Ki值均在188至22μM范围内。对应于多蛋白裂解位点P1'-P5'区域的肽在1mM浓度下对酶活性没有影响。肽抑制剂与 2 型登革热病毒 NS2B(H)-NS3p 复合物的同源模型的分子对接数据表明，非引物位点产物抑制剂的结合与相应底物的基态结合相似。 (c) 2005 Elsevier Inc. 保留所有权利。
The NS3 serine protease of dengue virus is required for the maturation of the viral polyprotein and consequently represents a promising target for the development of antiviral inhibitors. However, the substrate specificity of this enzyme has been characterized only to a limited extent. In this study, we have investigated product inhibition of the NS3 protease by synthetic peptides derived from the P6-P1 and the P1'-P5' regions of the natural polyprotein substrate. N-terminal cleavage site peptides corresponding to the P6-P1 region of the polyprotein were found to act as competitive inhibitors of the enzyme with K-i values ranging from 67 to 12 mu M. The lowest K-i value was found for the peptide representing the NS2A/NS2B cleavage site, RTSKKR. Inhibition by this cleavage site sequence vias analyzed by using shorter peptides, SKKR, KKR, KR, AGRR, and GKR. With the exception of the peptide AGRR which did not inhibit the protease at a concentration of 1 mM, all other peptides displayed Ki values in the range from 188 to 22 mu M. Peptides corresponding to the P1'-P5' region of the polyprotein cleavage sites had no effect on enzymatic activity at a concentration of 1 mM. Molecular docking data of peptide inhibitors to a homology-based model of the dengue virus type 2 NS2B(H)-NS3p co-complex indicate that binding of the non-prime site product inhibitors is similar to ground-state binding of the corresponding substrates. (c) 2005 Elsevier Inc. All rights reserved.