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Determination of active site of lysine-specific cysteine proteinase (Lys-gingipain) by use of a Porphyromonas gingivalis plasmid system.


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タイトル: Determination of active site of lysine-specific cysteine proteinase (Lys-gingipain) by use of a Porphyromonas gingivalis plasmid system.
著者: Ishida, Yutaka / Hu, Jinping / Sakai, Eiko / Kadowaki, Tomoko / Yamamoto, Kenji / Tsukuba, Takayuki / Kato, Yuzo / Nakayama, Koji / Okamoto, Kuniaki
発行日: 2008年 6月
出版者: Elsevier Ltd
引用: Archives of oral biology, 53(6), pp.538-544; 2008
抄録: Porphyromonas gingivalis, a major etiological bacterium of periodontal diseases, produces a unique lysine-specific cysteine proteinase (Lys-gingipain, Kgp) implicated in the virulence of this organism. Our observations show the expression of a catalytically active recombinant Kgp in a P. gingivalis Kgp-null mutant and the restoration of its functions by the use of a shuttle plasmid vector stable in P. gingivalis. The Kgp-expressing mutant exhibited a similar catalytic activity to that of the wild-type strain. This mutant also restored the ability to form black-pigmented colonies on blood agar plates and to generate a 19-kDa haemoglobin receptor protein responsible for haemoglobin binding. In order to establish the importance of the active-site Cys residue and elucidate its role in bacterial black pigmentation we constructed three Kgp mutants with changed potential active-site Cys residues. The cells expressing a single mutation (C476A) showed the high Kgp activity and the black pigmentation. In contrast, the cells expressing the single mutant (C477A) and the double mutant (C476A/C477A) exhibited neither Kgp activity nor black pigmentation. These results indicate that the 477th Cys residue is essential for both the Kgp activity and the black pigmentation of P. gingivalis.
記述: 長崎大学学位論文 学位記番号:博(医歯薬)甲第139号 学位授与年月日:平成20年2月27日
キーワード: Arg-gingipain / Lys-gingipain / Periodontal diseases / Porphyromonas gingivalis / Recombinant expression
URI: http://hdl.handle.net/10069/23114
ISSN: 00039969
DOI: 10.1016/j.archoralbio.2008.01.004
PubMed ID: 18295742
関連リンク : http://hdl.handle.net/10069/16965
権利: Copyright © 2008 Elsevier Ltd All rights reserved.
資料タイプ: Journal Article
原稿種類: author
出現コレクション:040 学術雑誌論文

引用URI : http://hdl.handle.net/10069/23114

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