Oviductin, the Oviductal Protease That Mediates Gamete Interaction by Affecting the Vitelline Coat in Bufo japonicus: Its Molecular Cloning and Analyses of Expression and Posttranslational Activation
| Autor: | Hideo Kubo, Koichi Mita, Yasusi Sugimoto, Chiaki Katagiri, Kazufumi Takamune, Masateru Hiyoshi |
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| Rok vydání: | 2002 |
| Předmět: |
DNA
Complementary Vitelline Duct medicine.medical_treatment Molecular Sequence Data hormonally induced gene expression Oviducts Biology Molecular cloning oviductin cDNA oviductal pars recta Western blot Complementary DNA medicine Animals Bufo japonicus Amino Acid Sequence RNA Messenger Cloning Molecular Molecular Biology Ovum chemistry.chemical_classification Serine protease Protease Molecular mass medicine.diagnostic_test urogenital system Serine Endopeptidases posttranslational processing Cell Biology Molecular biology Bufonidae Amino acid Cell biology Open reading frame chemistry biology.protein Female Protein Processing Post-Translational Sequence Alignment Developmental Biology |
| Zdroj: | Developmental Biology. 243:176-184 |
| ISSN: | 0012-1606 |
| DOI: | 10.1006/dbio.2001.0558 |
| Popis: | Previous studies indicated that the acquisition of egg fertilizability during transit through the pars recta portion of the oviduct in Bufo japonicus is accompanied by hydrolytic conversion of the vitelline coat 40- to 52-kDa components to 39-kDa components induced by a 66-kDa serine protease, “oviductin.” In this study, we cloned a 3028-bp cDNA that contained an open reading frame encoding 974 amino acids with a calculated molecular mass of 107.6 kDa, including two protease domains and three repeats of CUB domains. Sequence analysis indicated that the catalytically active 66-kDa protein comprised an N-terminally located oviductin protease and two CUB domains. The oviductin gene was transcribed as a part of 6-kb mRNA that was expressed specifically in the cells lining the bottom of epithelial folds in the oviductal pars recta, and this expression was highly accelerated when the pars recta fragments were cultured in the presence of hCG. Western blot analyses using antibodies against a protease domain revealed that the catalytically inactive 102-kDa proteins in the pars recta granules yield 66-kDa catalytically active and 82- and 59-kDa inactive molecules. We propose that the oviductin translated as 107.6-kDa precursors are processed both N- and C-terminally to give rise to a 66-kDa active form comprising a serine protease and two CUB domains. |
| Databáze: | OpenAIRE |
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