title: Optimization of the multiple enzymatic activities of the hepatitis C virus NS3 protein.
abstract: The hepatitis C virus (HCV) nonstructural protein 3 (NS3) is known to possess multiple enzymatic activities. In addition to its well-characterized protease activity, HCV NS3 also has ATP hydrolase (ATPase) and nucleic acid unwinding (helicase) activities. We systematically studied the effect of common reagents on all three enzymatic activities with a view to improving assay sensitivity for compound screening and profiling. Inclusion of the detergent lauryl dimethylamine oxide (LDAO) improves protease and helicase activities significantly, allowing robust assays at much lower NS3 concentrations. These conditions enable a particularly sensitive protease assay that uses picomolar concentrations of NS3.
authors: InterMune, Brisbane, CA 94005, USA.
authors: Shawn
authors: S
authors: Misialek
authors: Ravi
authors: R
authors: Rajagopalan
authors: Sarah K
authors: SK
authors: Stevens
authors: Leonid
authors: L
authors: Beigelman
authors: Scott D
authors: SD
authors: Seiwert
authors: Karl
authors: K
authors: Kossen
mesh_terms: D003902:Detergents / Q000494:pharmacology; D004123:Dimethylamines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016174:Hepacivirus / Q000201:enzymology*; D007700:Kinetics; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D003902:Detergents; D004123:Dimethylamines; D004791:Enzyme Inhibitors; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C014518:dodecyldimethylamine oxide