1996 Fiscal Year Final Research Report Summary
PROTEASES INVOLVED IN REGULATION OF EGG-ENVELOPES DURING FERTILIZATION AND DEVELOPMENT
| Project/Area Number |
07458193
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KATAGIRI Chiaki GRADUATE SCHOOL OF SCIENCE,HOKKAIDO UNIVERSITY,PROFESSOR, 大学院・理学研究科, 教授 (90000827)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Masakane GRADUATE SCHOOL OF SCIENCE,HOKKAIDO UNIVERSITY,ASSOC.PROFESSOR, 大学院・理学研究科, 助教授 (30202378)
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| Project Period (FY) |
1995 – 1996
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| Keywords | VITELLINE ENVELOPE / SPERM-VE BINDING / OVIDUCAL PARS RECTA / TRYPSIN-LIKE PROTEASE / HATCHING ENZYME / METALLOPROTEASE / ASTACIN FAMILY / CUB-DOMAIN |
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| Research Abstract |
Functions and molecular entities were studied of proteases that participate in fertilization and embryonic hatching of anuran amphibians. (a) The eggs of Bufo japonicus become fertilizable during passage through the pars recta (PR) portion of oviduct, accompanied by a significant increase of sperm-binding to vitelline envelope (VE). The sperm binding to VE was found to be mediated by carbohydrate moieties of VE,which was exposed as a result of partial hydrolysis of VE by a trypsin-like protease secreted from PR epithelial cells. The method was developed to isolated this protease, and partial amino acid sequences were determined for its further molecular cloning. (b) Using a previously cloned cDNA,UVS.2, as a probe, a 1.8kb insert (XHE) was cloned from a cDNA library from the NF st.25 Xenopus laevis embryos. XHE was found to code for hatching enzyme, since it encoded 514 amino acids including both signal and propeptide sequences, predicted mature enzyme of 425 amino acids containing metalloprotease of astacin family and CUB-domains. The antibodies against bacterially expressed XHE inhibited the VE digesting activity of the hatching medium, and localized in immunocytochemical stainings the reacting molecules specifically on secretory granules of hatching gland cells. Characterization of the hatching enzyme using these antibodies as probe indicated that the 60kDa enzyme consists of 40kDa molecules of protease domain and other portion which may be involved in recognition and/or processing of the substrate VE.
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Research Products
(12 results)
