| Project/Area Number |
08457481
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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 |
Morphological basic dentistry
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
SUGINAKA Hidekazu HIROSHIMA UNIV.SCH.OF DENTISTRY,PROFESSOR, 歯学部, 教授 (70028736)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIWARA Tamaki HIROSHIMA UNIV.SCH.OF DENTISTRY,RESEARCH ASSOCIATE, 歯学部, 助手 (90274092)
SUGAI Motoyuki HIROSHIMA UNIV.SCH.OF DENTISTRY, ASSOCIATE PROFESSOR, 歯学部, 助教授 (10201568)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1997: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1996: ¥6,700,000 (Direct Cost: ¥6,700,000)
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| Keywords | BACTERIOLYTIC ENZYME / AUTOLYSIN / ATL / STAPHYLOCOCCUS AUREUS / 黄色ブドウ球菌 / LTA / PCG |
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| Research Abstract |
The aim of this project was to understand regulation mechanism of Staphylococcus aureus bacteriolytic enzyme. We first grew S.aureus in chemically defined medium and performed pulsc/chasc experiment of atl gene product by using radio-labeled methionine. According to the results of immunprecipitation using anti-ATL antibody, it was clear that ATL was secreted as 138 kDa protein, and thereafter processed to 115 kDa, 85 kDa, 62 kDa and 51 kDa proteins. These results showed that ATL undergo posttranslational processing after it is translocated the cell membrane. We also purified ail gene products associated with S.aureus cells, and compared them with those purifed from culture supernatant. Accordingly, it became apparent that 62 kDa and 51 kDa proteins purified from extract of cell-associated protein were 62 kDa N-acetylmuramyl-L-alanine amidase and 51 kDa endo-beta-N-acetylglucosaminidase. Furthermore, electron microscopical observation using anti-ATL antibody revealed that there are two types of ail gene products, the one extractable with 3M LiCl and the other nonextractable with 3M LiCl. Matured ATL such as 51 kDa glucosaminidase and 62 kDa amidase are predominat in LiCl-extractable fraction and 138 kDa protein was rich in LiCl-nonextractable fraction. These results sugest that atl gene products associate with some unknown factor after it is translocated, and undergo processing and are released into cupernatant. We investigated the factor which interact with ATL.but only found that it is not protein. Purification of the factor was unsuccessful. and its identity remains unknown.
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