Identification and analysis of hepatitis C virus NS3 helicase inhibitors using nucleic acid binding assays
dc.contributor.author | Mukherjee, Sourav | |
dc.contributor.author | Hanson, Alicia M. | |
dc.contributor.author | Shadrick, William R. | |
dc.contributor.author | Ndjomou, Jean | |
dc.contributor.author | Sweeney, Noreena L. | |
dc.contributor.author | Hernadez, John J. | |
dc.contributor.author | Bartczak, Diana | |
dc.contributor.author | Li, Kelin | |
dc.contributor.author | Frankowski, Kevin J. | |
dc.contributor.author | Heck, Julie A. | |
dc.contributor.author | Arnold, Leggy A. | |
dc.contributor.author | Schoenen, Frank J. | |
dc.contributor.author | Frick, David N. | |
dc.date.accessioned | 2014-04-11T19:43:17Z | |
dc.date.available | 2014-04-11T19:43:17Z | |
dc.date.issued | 2012-06-27 | |
dc.identifier.citation | Mukherjee, Sourav, Alicia M Hanson, William R Shadrick, Jean Ndjomou, Noreena L Sweeney, John J Hernandez, Diana Bartczak, et al. 2012. “Identification and Analysis of Hepatitis C Virus NS3 Helicase Inhibitors Using Nucleic Acid Binding Assays.” Nucleic Acids Research 40 (17): 8607–21. http://dx.doi.org/10.1093/nar/gks623. | |
dc.identifier.uri | http://hdl.handle.net/1808/13459 | |
dc.description.abstract | Typical assays used to discover and analyze small molecules that inhibit the hepatitis C virus (HCV) NS3 helicase yield few hits and are often confounded by compound interference. Oligonucleotide binding assays are examined here as an alternative. After comparing fluorescence polarization (FP), homogeneous time-resolved fluorescence (HTRF®; Cisbio) and AlphaScreen® (Perkin Elmer) assays, an FP-based assay was chosen to screen Sigma’s Library of Pharmacologically Active Compounds (LOPAC) for compounds that inhibit NS3-DNA complex formation. Four LOPAC compounds inhibited the FP-based assay: aurintricarboxylic acid (ATA) (IC50 = 1.4 μM), suramin sodium salt (IC50 = 3.6 μM), NF 023 hydrate (IC50 = 6.2 μM) and tyrphostin AG 538 (IC50 = 3.6 μM). All but AG 538 inhibited helicase-catalyzed strand separation, and all but NF 023 inhibited replication of subgenomic HCV replicons. A counterscreen using Escherichia coli single-stranded DNA binding protein (SSB) revealed that none of the new HCV helicase inhibitors were specific for NS3h. However, when the SSB-based assay was used to analyze derivatives of another non-specific helicase inhibitor, the main component of the dye primuline, it revealed that some primuline derivatives (e.g. PubChem CID50930730) are up to 30-fold more specific for HCV NS3h than similarly potent HCV helicase inhibitors. | |
dc.description.sponsorship | National Institutes of Health [RO1 AI088001]; Research Growth Initiative Award [101X219] from the University of Wisconsin-Milwaukee Research Foundation; National Institutes of Health Molecular Libraries Initiative [U54 HG005031]. Funding for open access charge: University of Wisconsin-Milwaukee Research Foundation. | |
dc.publisher | Oxford University Press | |
dc.rights | This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. | |
dc.rights.uri | http://creativecommons.org/licenses/by-nc/3.0 | |
dc.title | Identification and analysis of hepatitis C virus NS3 helicase inhibitors using nucleic acid binding assays | |
dc.type | Article | |
kusw.kuauthor | Li, Kelin | |
kusw.kuauthor | Frankowski, Kevin J. | |
kusw.kuauthor | Schoenen, Frank J. | |
kusw.kudepartment | Department of Chemisty | |
kusw.oastatus | fullparticipation | |
dc.identifier.doi | 10.1093/nar/gks623 | |
kusw.oaversion | Scholarly/refereed, publisher version | |
kusw.oapolicy | This item meets KU Open Access policy criteria. | |
dc.rights.accessrights | openAccess |
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