| Project/Area Number |
16590363
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | KAGAWA UNIVERSITY |
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Principal Investigator |
MATSUSHITA Osamu Kagawa University, Faculty of Medicine, Molecular Microbiology, Assistant Professor, 医学部, 助教授 (00209537)
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| Co-Investigator(Kenkyū-buntansha) |
KOIDE Takaki Niigata University of Pharmacy and Appllied Life Sciences, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助教授 (70322253)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | clostridial collagenases / collagen-binding domain / NMR analysis / substrate recognition |
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| Research Abstract |
Three distinct domains [metallopeptidase catalytic domain (CD), PKD domain (PKD), and collagen-binding domain (CBD)] are present in clostridial collagenases. The variation in the number of PKDs and CBDs found in Clostridiun histolyticum collagenases class I (CD+PKD+2CBDs) and class II (CD+2PKDs+CBD) led us to investigate their evolutionary histories. In this study, facilitated by a novel purification method and universal PCR primers, we obtained six new sequences and carried out a phylogenetic analysis.
Sequences homologous to clostridial collagenases were divided into two subfamilies, M9A and M9B. The M9A sequences were found in proteobacteria such as Burkholderia and Vibrio with one exception Streptomyces coelicolor of Actinobacteria, while the M9B sequences were in the firmicutes Clostridium and Bacillus. S.coelicolor might have acquired the M9A enzyme by horizontal gene transfer. The M9A enzymes possess PPC domains (PPC) instead of CBDs at their C-termini. Although PPCs showed sequence similarity with CBDs, the calcium-binding and substrate-binding sites identified in CBDs were not conserved in the PPCs. CBD may have evolved from PPCs by acquiring these sites. Among M9B enzymes, the numbers of PKDs and CBDs varied from none to two and one to three, respectively.
Inference on the loss and gain of PKDs and CBDs, carried out by taking into account the phylogenetic relationships of the clostridial species, indicated their frequent loss and gain in Clostridial lineages.
Clostridia may have modulated the number of C-terminal domains frequently to adapt themselves to their specific pathogenic/saprophytic life styles.
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