Abstract

We previously identified the antibody-binding site of a monoclonal antibody (mAb 79.0) on varicella-zoster virus (VZV) glycoprotein I (gpI) and showed that this monoclonal antibody binds to both VZV gpI and gpIV (Vafai et al., J. Virol. 62, 2544, 1988). In this study, a synthetic peptide comprising the mAb 79.0 binding site (designated e1) was prepared and anti-peptide antibodies (RAnti-e1) were raised in rabbit. RAnti-e1 recognized the primary translation products encoded by VZV genes 67 (gpIV) and 68 (gpI). To further localize the binding site of RAnti-e1 on VZV gpIV, the gpIV gene cloned in pGEM transcription vector was cleaved at different locations to generate four truncated DNA fragments. RNA was transcribed from each truncated gpIV fragment, translated in vitro and immunoprecipitated with RAnti-e1. The results indicated that RAnti-e1 binds an antigenic determinant within the first 153 amino acid residues on the primary translation product of VZV gpIV. In addition, RAnti-e1 recognized the high-mannose intermediate but not the mature form of gpI in the infected cells or the translation products of gpIV glycosylated in vitro in the presence of canine microsomal membrane. These results: (a) confirmed the existence of a shared antigenic determinant on both VZV gpI and gpIV; and (b) indicated that the addition of terminal sugar modification may influence the conformation of gpI and gpIV with respect to the antigenic determinant recognized by RAnti-e1.

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