Mapping of epitopes on porcine zona pellucida-3 α by monoclonal antibodies inhibiting oocyte-sperm interaction

Gupta, S. K. ; Chadha, K. ; Harris, J. D. ; Yurewicz, E. C. ; Sacco, A. G. ; Kolluri, S. K. ; Afzalpurka, A. (1996) Mapping of epitopes on porcine zona pellucida-3 α by monoclonal antibodies inhibiting oocyte-sperm interaction Biology of Reproduction, 55 (2). pp. 410-415. ISSN 0006-3363

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Official URL: http://www.biolreprod.org/content/55/2/410.short

Related URL: http://dx.doi.org/10.1095/biolreprod55.2.410

Abstract

Zona pellucida glycoprotein 3 α(ZP3 α) has been designated as the primary sperm receptor ligand in porcine gamete interaction. In this study, epitopes were mapped on porcine ZP3 α (pZP3 α) by using monoclonal antibodies (mAbs) possessing in vitro contraceptive efficacy. Using Western blots, we tested recombinant pZP3 α fragments expressed as fusion proteins in Escherichia coli and corresponding to pZP3 α precursor protein amino acid residues 18-142 (F1), 140-243 (F2), 239-363 (F3), and 359-462 (F4), for reactivities with mAbs. MAb-403 reacted with F3, and mAbs-412 and -421 with F2. MAb-420 showed weak reactivity with F1. Synthesis of overlapping 12-mer peptides further resolved the epitope for mAb-420 to amino acid residues 133-144, mAb-421 to 157-168, mAb-412 to 205-216, and mAb-403 to 301-312. MAbs-412 and -420 inhibited the binding of boar sperm to zona-encased porcine oocytes. These results, the first to define peptide epitopes of pZP3 α, should assist in the design of a synthetic peptide-based immunocontraceptive vaccine.

Item Type:Article
Source:Copyright of this article belongs to Society for the Study of Reproduction.
ID Code:72153
Deposited On:28 Nov 2011 06:09
Last Modified:28 Nov 2011 06:09

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