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Regular Article |
a Graduate Institute of Life Science,
b Department of Biology and Anatomy,
c Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan 114, Republic of China
d Division of Urology, Department of Surgery, Tri-Service General Hospital, Taipei, Taiwan 114, Republic of China
e Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan 115, Republic of China
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
-form of 3451 base pairs (bp) and ß-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.
First decision: 18 April 2001.
1 This study was supported in part by grants from the National Science Council, Republic of China (NSC 85-2331-B016-097 and NSC 86-2314-B016-064).
2 Correspondence: Hwan-Wun Liu, Institute of Preventive Medicine, National Defense Medical Center, 161, Sect. 6, Minchuan E. Rd., Taipei, Taiwan 114, ROC. FAX: 886 2 87923159; hwanliu{at}ndmctsgh.edu.tw
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