Peptide inhibitor of Japanese encephalitis virus infection targeting envelope protein domain III

• Specific JEV E DIII-interacting peptides were successfully selected using a phage display library.
• A highly enriched peptide named P3 strongly inhibited JEV.
• P3 was presumed to block JEV attachment by binding to the N terminus of E DIII.

Japanese encephalitis virus (JEV) is a major cause of acute viral encephalitis in both humans and animals. Domain III of the virus envelope glycoprotein (E DIII) plays an important role in the interaction of viral particles with host cell receptors to facilitate viral entry. Intervention of the interaction between E DIII and its cognate host cell receptor would provide an important avenue for inhibiting JEV infection. A phage display peptide library was therefore panned against E DIII, which resulted in the identification of several peptides. One peptide, named P3, inhibited JEV infection of BHK-21 cells with an IC50 of ∼1 μM and an IC90 at ∼100 μM. Further characterization revealed that P3 bound to E DIII with a Kd of 6.06 × 10−6 M and inhibited JEV infection by interfering with viral attachment to cells. Based on in silico prediction by ZDOCK, P3 was found to interact with E DIII via a hydrophobic pocket, which was confirmed by the binding assay of P3 to the V357A mutant. P3 was hypothesized to bind to E DIII by interacting with the sties adjacent to the BC and DE loops, which might interfere with the binding of JEV to cellular receptors, thus impeding viral infection. This newly isolated peptide may represent a new therapeutic candidate for treatment of JEV.