A FRET-based assay for the discovery of West Nile Virus NS2B-NS3 protease inhibitors
A FRET-based assay for the discovery of West Nile Virus NS2B-NS3 protease inhibitors
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DOI：
10.1016/j.bmcl.2013.06.081
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发表时间：
2013-09-01
影响因子：
2.7
通讯作者：
Salzameda, Nicholas T.
中科院分区：
文献类型：
--
作者：
Adamek, Rebecca N.;Maniquis, Roxanne V.;Salzameda, Nicholas T.
西尼罗河病毒（WNV）自20世纪90年代初以来一直是世界性的流行病。目前没有治疗西尼罗河病毒感染的治疗方法。一种特殊的治疗途径是抑制NS 2B-NS 3蛋白酶，这是一种对WNV复制至关重要的酶。为了增加NS 2B-NS 3蛋白酶抑制剂的数量，我们报告了一种新的基于FRET的高通量检测方法，用于发现WNV NS 2B-NS 3蛋白酶抑制剂。对于该测定，合成基于FRET的肽底物，并用NS 2B-NS 3蛋白酶进行动力学表征。新底物的Km为3.35 +/- 0.31 μ M，k（cat）为0.0717 +/- 0.0016 s（-1），k（cat）/K-m为21，400 +/- 2000 M-1 s（-1）。(C)2013爱思唯尔有限公司保留所有权利。
The West Nile Virus (WNV) has been a worldwide epidemic since the early 1990s. Currently there are no therapeutic treatments for WNV infections. One particular avenue of treatment is inhibition of the NS2B-NS3 protease, an enzyme that is crucial for WNV replication. In our effort to increase the number of NS2B-NS3 protease inhibitors, we report a novel FRET-based high throughput assay for the discovery of WNV NS2B-NS3 protease inhibitors. For this assay, a FRET-based peptide substrate was synthesized and kinetically characterized with the NS2B-NS3 protease. The new substrate exhibits a K-m of 3.35 +/- 0.31 mu M, a k(cat) of 0.0717 +/- 0.0016 s(-1) and a k(cat)/K-m of 21,400 +/- 2000 M-1 s(-1). (C) 2013 Elsevier Ltd. All rights reserved.