Fertilization antigen-1: cDNA cloning, testis-specific expression, and immunocontraceptive effects.

Abstract

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.

• Authors: X Zhu, R K Naz
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 94 Issue 9 Pg. 4704-9 (Apr 29 1997) ISSN: 0027-8424 [Print] United States
• PMID: 9114055 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antibodies
• Antigens
• DNA, Complementary
• Glycoproteins
• Recombinant Proteins
• fertilization antigen

Topics

• Amino Acid Sequence
• Animals
• Antibodies (pharmacology)
• Antigens (genetics, immunology, isolation & purification)
• Base Sequence
• Cloning, Molecular
• Contraception, Immunologic
• DNA, Complementary (genetics)
• Female
• Glycoproteins (genetics, immunology, isolation & purification)
• Male
• Mice
• Molecular Sequence Data
• Recombinant Proteins (immunology, isolation & purification)
• Sequence Analysis, DNA
• Sperm-Ovum Interactions (immunology)
• Spermatozoa
• Testis (physiology)
• Tissue Distribution
• Zona Pellucida