Fine mapping of sequential neutralization epitopes on the subunit protein VP8 of human rotavirus

Jennifer Kovacs-Nolan, Dongwan Yoo, Yoshinori Mine

Research output: Contribution to journalArticlepeer-review

Abstract

The epitopes of the HRV (human rotavirus), especially those involved in virus neutralization, have not been determined in their entirety, and would have significant implications for HRV vaccine development. In the present study, we report on the epitope mapping and identification of sequential neutralization epitopes, on the Wa strain HRV subunit protein VP8, using synthetic overlapping peptides. Polyclonal antibodies against recombinant Wa VP8 were produced previously in chicken, and purified from egg yolk, which showed neutralizing activity against HRV in vitro. Overlapping VP8 peptide fragments were synthesized and probed with the anti-VP8 antibodies, revealing five sequential epitopes on VP8. Further analysis suggested that three of the five epitopes detected, M1-L10, I55-D66 and L223-P234, were involved in virus neutralization, indicating that sequential epitopes may also be important for the HRV neutralization. The interactions of the antibodies with the five epitopes were characterized by an examination of the critical amino acids involved in antibody binding. Epitopes comprised primarily of hydrophobic amino acid residues, followed by polar and charged residues. The more critical amino acids appeared to be located near the centre of the epitopes, with proline, isoleucine, serine, glutamine and arginine playing an important role in the binding of antibody to the VP8 epitopes.

Original languageEnglish (US)
Pages (from-to)269-275
Number of pages7
JournalBiochemical Journal
Volume376
Issue number1
DOIs
StatePublished - Nov 15 2003
Externally publishedYes

Keywords

  • Epitope characterization
  • Immunoglobulin Y
  • Peptide synthesis
  • VP8
  • Virus neutralization
  • Wa strain

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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