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

Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4704-9. doi: 10.1073/pnas.94.9.4704.

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

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

Substances

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

Associated data

  • GENBANK/U95114