| Project/Area Number |
16591832
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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 |
Morphological basic dentistry
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
KAMAGUCHI Arihide Dept. of Oral Microbiology, School of Dentistry, Health Sciences Univ. of Hokkaido, Associated Professor, 歯学部, 助教授 (40133235)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Porphyromonas gingivalis / autolysis / cell death / stringent response / Rgp / oral bacteria / biofilm / relA / spoT / Prophyromonas gingivalis / growth / peptide / medium / Stringent response / relA gene / spot gene / ppk gene / Program death / rpoS gene |
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| Research Abstract |
Porphyromonas gingivalis is a putative major etiology agent of adult periodontitis. It is suppose that P. gingivalis may survive in the oral biofilm. However, P. gingivalis quickly autolyzes in the culture media. To examine the molecular components of the P gingivalis autolysate, two-dimensional gel electrophoresis was performed. Many protein spots varied both in number and volume. One of these spots included Rgp as determined by N-terminal amino acid sequencing. Mutant of N-acetylmuramoyl-L-alanine amidase like protein gene of P. gingivalis was constructed by alleic exchange. Autolysis rate of the autolysin mutant was the same as that of parent strain. These results indicate that many autolysin may act the autolysis of P. gingivalis.
From consideration that inside of oral biofilm may be innutritious space, mimic condition of oral biofilm was prepared by low concentration of THM medium. Major component of THM medium was Trypyone which is composed from peptide. P gingivalis could growth with metabolizing not amino acid but peptide. In low concentration of THM medium, autolysis rate of P gingivalis was decreased and living cells was increased. Proteome analysis of P. gingivalis cells in the low concentration of THM medium indicated that expression of many proteins decreased. It is considered that low concentration of THM medium is stringent state for P. gingivalis. One of the stringent response gene (relA/spot) mutant of P gingivalis decreased autolysis rate than parent strain. It was also observed that many protein expression were differ from relA/spot mutant and parent strain.
These results suggest that high peptide concentration induce autolysis (programmed cell death) and low peptide concentration in vitro induce the stringent response gene expression, consequently autolysis rate of P. gingivalis turned slow. This mechanism may be possible to explain the survival of P. gingival in biofilm.
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