2004 Fiscal Year Final Research Report Summary
Establishment of protein expression system in Porphyromonasi gingivalis
| Project/Area Number |
15591971
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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 |
Functional basic dentistry
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| Research Institution | NAGASAKI UNIVERSITY |
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Principal Investigator |
OKAMOTO Kuniaki Nagasaki University, Graduate School and Dental Sciences, Associate Professor, 大学院・医歯薬学総合研究科, 助教授 (10311846)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Yuuzo Nagasaki University, Graduate School and Dental Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (20014128)
SHIBATA Mitsue Nagasaki University, Graduate School and Dental Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (20274665)
SAKAI Eiko Nagasaki University, Graduate School and Dental Sciences, Research Associate, 大学院・医歯薬学総合研究科, 助手 (10176612)
NAKAYAMA Koji Nagasaki University, Graduate School and Dental Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (80150473)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Arg-gingipain / cysteine proteinase / Lys-gingipain / periodontitis disease / Porphysomonas gingivalis / transport |
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| Research Abstract |
Porphyromonas gingivalis(P.gingivalis) is a Gram-negative anaerobic bacterium and produces a number of proteinsaes. Among them, Arginine-specific cysteine proteinase(Arg-gingipain, Rgp) and lysine-specific cysteine proteinase(Lys-gingipain, Kgp) are implicated as major virulence factors in the development and progression of chronic periodontaitis. Rgp is encoded by two separate rgp genes (rgpA and rgpB), whereas Kgp is encoded by the single gene (kgp). The initial translation products of rgpA and kgp genes primarily consist of two functional parts : proteinase domain and adhesin domain. Furthermore, the C-terminal adhesin domains are derived into three or four subdomains (HGP44,HGP15,HGP17 and HGP27 for RgpA ; nHGP44,HGP15 and cHGP44 for Kgp). These C-terminal adhesin domains are highly homologous. In this study, we focused Kgp and established its expression system. Furthermore, we attempted to decide the active sites for Kgp. Although the active site of Rgp have been decided, that of Kgp was not clear due to have two cysteine residues in putative active site. Therefore, we introduced point mutation in the two putative active sites of Kgp. First, we constructed a plasmid that 248 cysteine (^<248>Cys) and 249 cysteine (^<249>Cys) were exchanged with alanine and introduced into Kgp-deficient strain KDP129. The mutation of both sites resulted in lack of emzymatic activity of Kgp, although the expression of Kgp was recognized by Western blot analysis. Next, we constructed a plasmid that point mutation plasmid against each active site and introduced into KDP129. These two mutants had the emzymatic activity of Kgp. These results suggested that ^<248>Cys and ^<249>Cys residues were important for the activity of Kgp.
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[Journal Article] Laminin □2 Is Essential for Odontoblast Differentiation Regulation Dentin Siaroprotein Expression.2004
Author(s)
Yuasa K, Fukumoto S, Kamasaki Y, Yamada A, Fukumoto E, Kanaoka K, Saito K, Harada H, Arikawa-Hirasawa E, Miyagoe-Suzuki Y, Takeda S, Okamoto K, Kato Y, Fujiwara T.
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Journal Title
J.Biol.Chem. 279
Pages: 10286-10292
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Roles of Arg- and Lys-gingipains in Coaggregation of Porphyromonas gingivalis : Identification of Its Responsible Molecules in Translation Products tf rgpA, kgp and HagA Genes.2004
Author(s)
Abe N, Baba A, Takii R, Nakayama K, Kamaguchi A, Shibata Y, Abiko Y, Okamoto K, Kadowaki T, Yamamoto K.
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Journal Title
Biol.Chem. 385
Pages: 1041-1047
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Identification of a new membrane-associated protein which influences transport/maturation of gingipains and adhesions of Porphyromonas gingivalis.2004
Author(s)
Sato k., Sakai E., Veith P.D., Shoji M., Kikuchi Y., Yukitake H., Ohara N., Naito M., Okamoto K., Reynolds E.G., Nakayama K.
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Journal Title
J.Biol.Chem. 280
Pages: 8668-8677
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Association of cathepsin E deficiency with development of atopic dermatitis.2003
Author(s)
Tsukuba T., Okamoto K., Okamoto Y., Yanagawa M., Kohmura K., Yasuda Y., Uchi H., Nakahara T., Furue M., Nakayama K., Kadowaki T., Yamamoto K., Nakayama K-I.
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Journal Title
J.Biochem. 134
Pages: 893-902
Description
「研究成果報告書概要(欧文)」より
