Studies on iron acquisition mechanism of periodontal pathogen
Project/Area Number |
09671877
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Morphological basic dentistry
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Research Institution | Matsumoto Dental University, School of Dentistry |
Principal Investigator |
FUJIMURA Setsuo Matsumoto Dental University, assoc.prof., 歯学部, 助教授 (40045505)
|
Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Takeshi Matsumoto Dental University, prof., 歯学部, 教授 (60064656)
|
Project Period (FY) |
1997 – 1998
|
Project Status |
Completed (Fiscal Year 1998)
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Budget Amount *help |
¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
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Keywords | p.gingivalis / periodontopathogen / iron acquisition / proteinases / プロテイナーゼ |
Research Abstract |
Two arginine specific proteinases (RGP-A, RGP-B) and a lysine specific proteinase (KGP) were purified from the solubilized materials by a detergent of the envelope of Porphyromonas gingivalis ATCC 33277. The three enzymes were activated by reducing reagents. RGP-B was remarkably activated by glycyiglycine. Both RGP-A and RGP-B were inhibited by PCMB, NEM, EDTA, luepeptin, E64, and antipain. KGP was inhibited by PCMB, NEM, and TLCK.Moecular masses of RGP-A and RGP-B were 43 kDa and that of KGP was 48 kDa. Hydrolytic activities of RGP and KGP to the synthetic chromogenic substrates were limited to those containing arginine and lysine at the P-ipositions, respectively. Hemoglobin was almost completely digested when it was treated by RGP-B.Furthermore, it was found that a fraction of the hydrolytic products separated by Sepliadex G-100 gel filtration stimulated the growth ofresting cells of P.gingivlis derived by heme- depleted condition, indicating RGP-B plays a partial role to provide iron source from hemoglobin. P gingivalis cells expressed a prominent 19 kDa protein when grown on blood agar plates. Analysis of N-terminal amino acid sequence indicated that 19 kDa protein was encoded by HGP15 of RGP.The HGP 15 domain protein was purified from an HGP15-overproducing E.coli and was found that to have the ability to bind hemoglobin in a pH-dependent manner. The anti-HGP15 antiserum reacted with the 19 kDa hemoglobin binding protein. P.gingivalis wild-type strain showed pH-dependent hemoglobin adsorption, whereas its non-pigmented mutants that produced no HGPI5-related proteins showed deficiency in hemoglobin adsorption. These results indicate a close relationship among GPI5 production, adsorption, and heme accumulation of P gingivalis.
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Report
(3 results)
Research Products
(10 results)