A method to simultaneously monitor hepatitis C virus NS3 helicase and protease activities.
A method to simultaneously monitor hepatitis C virus NS3 helicase and protease activities.
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DOI：
10.1007/978-1-60327-355-8_16
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发表时间：
2010
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中科院分区：
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丙型肝炎病毒NS 3蛋白含有N-末端丝氨酸蛋白酶和C-末端解旋酶，解旋RNA或DNA双链体。HCV NS 3蛋白是临床试验中几种抗病毒药物的靶标，其抑制蛋白酶功能。报道了使用荧光光谱法和单链重组蛋白在单一反应中同时监测NS 3蛋白的解旋酶和蛋白酶功能的方法，其中NS 3与其蛋白酶激活剂NS 4A融合。所述方法真实的时间一起监测两种活性，并且适合于高通量筛选。这种新方法可用于鉴定抑制NS 3的解旋酶和蛋白酶活性的化合物。
The hepatitis C virus NS3 protein contains a N-terminal serine protease and a C-terminal helicase that unwinds RNA or DNA duplexes. The HCV NS3 protein is the target for several antiviral drugs in clinical trials, which inhibit the protease function. A method is reported to simultaneously monitor the helicase and protease function of the NS3 protein in a single reaction using fluorescence spectroscopy and a single chain recombinant protein where NS3 is fused to its protease activator NS4A. The method monitors both activities together in real time and is amenable to high throughput screening. This new procedure could be used to identify compounds that inhibit both the helicase and protease activity of NS3.