Search Results
You are looking at 1 - 2 of 2 items for
- Author: K. Koyama x
- Refine by access: All content x
Search for other papers by A. Hasegawa in
Google Scholar
PubMed
Search for other papers by K. Koyama in
Google Scholar
PubMed
Search for other papers by Y. Okazaki in
Google Scholar
PubMed
Search for other papers by M. Sugimoto in
Google Scholar
PubMed
Search for other papers by S. Isojima in
Google Scholar
PubMed
Mammalian zona pellucida is an attractive target for developing a contraceptive vaccine. In this study, the amino acid sequence of a core protein of porcine zona pellucida glycoprotein ZP4 was determined by peptide mapping and cDNA cloning. Two kinds of ZP4 peptides were identified: one consisted of 128 amino acid residues and the other of 133 amino acid residues with an additional five amino acid sequence at the carboxy-terminal end of the 128 amino acid peptide. Both peptides had two potential N-linked glycosylation sites. The 128 amino acid peptide showed 39.1% similarity to the amino-terminal region of mouse ZP2 polypeptide. The positions of five cysteine residues were the same for porcine ZP4 and mouse ZP2. The cloned cDNA possessed an additional 195 nucleotides at the 3' end of the sequence corresponding to the 133 amino acid peptide. This additional sequence was found to encode the amino-terminal 10 amino acid sequence of porcine ZP2 polypeptide. These results suggest that porcine ZP4 and ZP2 are derived from a common parent polypeptide by proteolytic cleavage at the position between 133 and 134 residues.
Search for other papers by A. Hasegawa in
Google Scholar
PubMed
Search for other papers by N. Yamasaki in
Google Scholar
PubMed
Search for other papers by M. Inoue in
Google Scholar
PubMed
Search for other papers by K. Koyama in
Google Scholar
PubMed
Search for other papers by S. Isojima in
Google Scholar
PubMed
The zona pellucida glycoprotein that surrounds the mammalian oocyte has several target antigens that have potential use in the development of a contraceptive vaccine. In the present study, an epitope sequence recognized by a monoclonal antibody to the porcine zona pellucida glycoprotein ZP4 was determined. Three candidate peptides were synthesized, based on an epitope mapping by cDNA and an analysis of chain flexibility of porcine ZP4. Only one synthetic peptide, corresponding to amino acid positions 50–67, reacted with the monoclonal antibody; the other synthetic peptides, corresponding to positions 60–79 and 70–100, did not react. The reactive epitope was identified as CTYVLDPENL, corresponding to positions 50–59 of porcine ZP4. The peptide inhibited the reaction of the monoclonal antibody binding to native ZP4 in a dose-dependent manner. When the synthetic peptide 50–67 was used to immunize mice, the resultant antisera reacted not only with the synthetic peptide but also with native pig zona pellucida. In addition, anti-peptide 50–67 antibody inhibited porcine fertilization in vitro. It is thus concluded that the peptide identified as an epitope for the monoclonal antibody would be a promising candidate for the development of a contraceptive vaccine.
