Mapping of epitopes relevant for induction of acrosome reaction on human zona pellucida glycoprotein-4 using monoclonal antibodies.

Wan-Xiang Xu, Beena Bhandari, Ya-Ping He, Hai-Ping Tang, Sanchita Chaudhary, Pankaj Talwar, Satish K Gupta, Jian Wang

Department of Reproductive Biology, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.

American journal of reproductive immunology (New York, N.Y. : 1989) 2012 Dec

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To decipher structural and functional aspects of human zona pellucida glycoprotein-4 (ZP4), the epitopes recognized by monoclonal antibodies (MAbs) have been mapped. Recombinant human ZP4-mediated induction of acrosome reaction in human sperm was studied in the absence and presence of ZP4-specific MAbs. The epitopes of MAbs were mapped using recombinant peptides expressed in Escherichia coli. Monoclonal antibodies (MA-1662, MA-1671) against human ZP4 showed specific binding to ZP matrix of human eggs in an indirect immunofluorescence assay. Both the antibodies showed significant (P < 0.05) inhibition in the baculovirus-expressed recombinant ZP4-mediated acrosome reaction. MA-1671 recognized N-terminal fragment of ZP4 and minimal epitope mapped to amino acid residues 126-130 (PARDR), whereas MA-1662 reacted to C-terminal fragment and minimal epitope mapped to amino acid residues 256-260 (ENELV). The epitopes corresponding to both N- and C-terminal parts of human ZP4 may be relevant for its biological activity. © 2012 John Wiley & Sons A/S.

Citation

Wan-Xiang Xu, Beena Bhandari, Ya-Ping He, Hai-Ping Tang, Sanchita Chaudhary, Pankaj Talwar, Satish K Gupta, Jian Wang. Mapping of epitopes relevant for induction of acrosome reaction on human zona pellucida glycoprotein-4 using monoclonal antibodies. American journal of reproductive immunology (New York, N.Y. : 1989). 2012 Dec;68(6):465-75