A method to simultaneously monitor hepatitis C virus NS3 helicase and protease activities.
The hepatitis C virus NS3 protein contains an 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.
Frick
David N
DN
Department of Biochemistry & Molecular Biology, New York Medical College, Valhalla, NY, USA.
Ginzburg
Olya
O
Lam
Angela M I
AM
D000998:Antiviral Agents / Q000378:metabolism / Q000627:therapeutic use; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism; D006526:Hepatitis C / Q000188:drug therapy; D006801:Humans; D010447:Peptide Hydrolases / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000998:Antiviral Agents; D010447:Peptide Hydrolases; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases