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Abstract
Viruses manipulate the life cycle in host cells via the use of viral properties and host machineries. Development of antiviral peptides against dengue virus (DENV) infection has previously been concentrated on blocking the actions of viral structural proteins and enzymes in virus entry and viral RNA processing in host cells. In this study, we proposed DENV NS1, which is a multifunctional non-structural protein indispensable for virus production, as a new target for inhibition of DENV infection by specific peptides. We performed biopanning assays using a phage-displayed peptide library and identified 11 different sequences of 12-mer peptides binding to DENV NS1. In silico analyses of peptide-protein interactions revealed 4 peptides most likely to bind to DENV NS1 at specific positions and their association was analysed by surface plasmon resonance. Treatment of Huh7 cells with these 4 peptides conjugated with N-terminal fluorescent tag and C-terminal cell penetrating tag at varying time-of-addition post-DENV infection could inhibit the production of DENV-2 in a time- and dose-dependent manner. The inhibitory effects of the peptides were also observed in other virus serotypes (DENV-1 and DENV-4), but not in DENV-3. These findings indicate the potential application of peptides targeting DENV NS1 as antiviral agents against DENV infection.
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Details
1 Mahidol University, Division of Molecular Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
2 Rangsit University, Faculty of Pharmacy, Pathum Thani, Thailand (GRID:grid.412665.2) (ISNI:0000 0000 9427 298X); University of Siegen, Department of Chemistry and Biology, Siegen, Germany (GRID:grid.5836.8) (ISNI:0000 0001 2242 8751)
3 Mahidol University, Department of Anatomy, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
4 National Science and Technology Development Agency, Molecular Biology of Dengue and Flaviviruses Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology, Bangkok, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408); Mahidol University, Division of Dengue Hemorrhagic Fever Research, Research Department, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490); Mahidol University, Siriraj Center of Research Excellence in Dengue and Emerging Pathogens, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
5 Mahidol University, Department of Biochemistry, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand (GRID:grid.10223.32) (ISNI:0000 0004 1937 0490)
6 National Science and Technology Development Agency, Protein-Ligand Engineering and Molecular Biology Research Team, Medical Molecular Biotechnology Research Group, National Center for Genetic Engineering and Biotechnology, Pathum Thani, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408)
7 National Biobank of Thailand, National Science and Technology Development Agency, Pathum Thani, Thailand (GRID:grid.425537.2) (ISNI:0000 0001 2191 4408)