Fertilization antigen-1: cDNA cloning, testis-specific expression, and immunocontraceptive effects.
cDNA encoding for a sperm antigen, designated fertilization antigen (FA-1), was cloned and sequenced from murine testis cDNA-lambdagt11 expression library using FA-1 mAb. Computer-generated translation analysis of 649-bp cDNA yielded an ORF of 164 amino acids with the first ATG Met start codon at nucleotide 81 and the stop codon TAA at nucleotide 577 and a polyadenylylation tail following the stop codon. The translated protein has a calculated molecular mass of 18.2 kDa and a potential N-linked glycosylation site at amino acids 158-160, besides at least two O-linked glycosylation sites. The hydropathy plot generated from the deduced amino acid sequence indicated it to be a membrane-anchored peptide. Extensive computer search in the GenBank, National Biomedical Research Foundation, and Swiss sequence banks did not identify any known nucleotide/amino acid sequence having homology with FA-1 cDNA or deduced amino acids, indicating it to be a novel protein. Northern blot analysis and reverse transcription-PCR indicated testis-specific expression of FA-1 antigen. The FA-1 cDNA was subcloned into pGEX-2T vector and expressed in glutathione S-transferase gene fusion system to obtain the recombinant protein. The recombinant protein specifically reacted with ZP3 of oocyte zona pellucida and its affinity-purified antibodies completely blocked sperm-zona pellucida interaction in mice. These findings suggest that the sperm-specific recombinant FA-1 antigen is an attractive candidate for the development of a contraceptive vaccine.
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Division of Research, Department of Obstetrics and Gynecology, Medical College of Ohio, Toledo, OH 43699, USA.
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D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D000941:Antigens / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D003269:Contraception, Immunologic; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D013084:Sperm-Ovum Interactions / Q000276:immunology*; D013094:Spermatozoa; D013737:Testis / Q000502:physiology*; D014018:Tissue Distribution; D015044:Zona Pellucida
D000906:Antibodies; D000941:Antigens; D018076:DNA, Complementary; D006023:Glycoproteins; D011994:Recombinant Proteins; C054151:fertilization antigen