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1 September 2001 Cloning and Characterization of a Complementary DNA Encoding a Human Epididymis-Associated Disintegrin and Metalloprotease 7 Protein
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Abstract

Mammalian spermatozoa interact with the proteins secreted by the epididymis to develop fertility. Transmembrane proteins that possess a disintegrin and metalloprotease (ADAM) domains are shown to be closely related to spermatogenesis and fertilization. Our previous study demonstrated that GP-83, a glycoprotein secreted by the epididymis, was conjugated to mature sperm. In this study, a 2.1-kilobase (kb) GP-83-expressing insert was isolated from a cDNA library of human epididymis by immunoscreening using GP-83-specific antiserum. The 5′ end rapid amplification of cDNA ends (RACE) and 3′-RACE of the 2.1-kb insert elucidated two isoforms of GP-83-encoding cDNA sequences, an α-form of 3451 base pairs (bp) and β-form of 2643 bp. Both forms exhibit the same open reading frame of 2262 bp predicting a peptide of 754 amino acid residues. Deduced amino acid sequence revealed signal sequence, prodomain, metalloproteinase, disintegrin, cysteine-rich, epidermal growth factor-like, transmembrane, and cytoplasmic domains. The GP-83-encoding sequence was recognized as human ADAM7 due to significant homology to other ADAM7s. According to the DNA sequences elucidated in the Human Genome Project, h-ADAM7 was located at chromosome 8p22. Ex vivo expression confirmed that h-ADAM7 cDNA did encode GP-83. Northern blot analysis revealed two transcripts of 4 kb and 3 kb in the epididymis, but not in testis or other major tissues. These results indicate that the GP-83-encoding gene is a human epididymis-associated ADAM7 gene (human ADAM7, h-ADAM7) and may be involved in the sperm-egg interaction.

Yu-Chi Lin, Guang-Huan Sun, Yu-May Lee, Yaw-Wen Guo, and Hwan-Wun Liu "Cloning and Characterization of a Complementary DNA Encoding a Human Epididymis-Associated Disintegrin and Metalloprotease 7 Protein," Biology of Reproduction 65(3), 944-950, (1 September 2001). https://doi.org/10.1095/biolreprod65.3.944
Received: 22 March 2001; Accepted: 1 May 2001; Published: 1 September 2001
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