Peptide inhibitors of hepatitis C virus core oligomerization and virus production.
Hepatitis C virus (HCV) nucleocapsid assembly requires dimerization of the core protein, an essential step in the formation of the virus particle. We developed a novel quantitative assay for monitoring this protein-protein interaction, with the goal of identifying inhibitors of core dimerization that might block HCV production in infected Huh-7.5 hepatoma cells. Two core-derived, 18-residue peptides were found that inhibited the dimerization of a fragment of core comprising residues 1-106 (core106) by 68 and 63%, respectively. A third, related 15-residue peptide displayed 50% inhibition, with an IC50 of 21.9 microM. This peptide was shown, by fluorescence polarization, to bind directly to core106 with a Kd of 1.9 microM and was displaced by the unlabelled peptide with an IC50 of 18.7 microM. When measured by surface plasmon resonance, the same peptide bound core169 with a Kd of 7.2 microM. When added to HCV-infected cells, each of the three peptides blocked release, but not replication, of infectious virus. When measured by real-time RT-PCR, the RNA levels were reduced by 7-fold. The 15-residue peptide had no effect on HIV propagation. Such inhibitors may constitute useful tools to investigate the role of core dimerization in the virus cycle.
Kota
S
S
The Scripps Research Institute - Florida, Department of Infectology, 130 Scripps Way, #3C1 Jupiter, FL 33458, USA.
Coito
C
C
The Scripps Research Institute - Florida, Department of Infectology, 130 Scripps Way, #3C1 Jupiter, FL 33458, USA.
Mousseau
G
G
The Scripps Research Institute - Florida, Department of Infectology, 130 Scripps Way, #3C1 Jupiter, FL 33458, USA.
Lavergne
J-P
JP
Institut de Biologie et Chimie des Protéines, UMR5086, CNRS-Université Lyon I, IFR128, 7 Passage du Vercors, 69367 Lyon Cedex 07, France.
Strosberg
A D
AD
The Scripps Research Institute - Florida, Department of Infectology, 130 Scripps Way, #3C1 Jupiter, FL 33458, USA.
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D019281:Dimerization; D006678:HIV / Q000187:drug effects; D016174:Hepacivirus / Q000187:drug effects*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D010455:Peptides / Q000494:pharmacology*; D011485:Protein Binding; D014758:Viral Core Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D010455:Peptides; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus