1996 Fiscal Year Annual Research Report

歯周病発症における病原性プロテアーゼの役割と作用機序

Research Project

Project/Area Number	08407059
Research Category	Grant-in-Aid for Scientific Research (A)
Research Institution	Kyushu University
Principal Investigator	山本健二九州大学, 歯学部, 教授 (40091326)
Co-Investigator(Kenkyū-buntansha)	筑波隆幸九州大学, 歯学部, 助手 (30264055) 中西博九州大学, 歯学部, 助手 (20155774)
Keywords	歯周病 / 歯周病原菌 / Porphyromonas gingivalis / 病原菌プロテアーゼ / Arg-gingipain / Lys-gingipain / カテプシン / 病原性発現機序
Research Abstract	歯周病の主たる病態は、歯肉上皮ならびに結合組織などの比較的表層に存在する歯周組織の破壊とそれに引き続いて起こる歯槽骨こうした歯周組織の破壊には口腔細菌ならびに宿主細胞の産生するプロテアーゼが中心的役割を果たしていると考えられている。しかし、歯周病発症における病原性プロテアーゼの機能や作用機序については依然として不明な点が多く残されている。今年度は歯周炎の主要な病原性細菌であるPorphyromonas gingivalisが産生する病原性プロテアーゼの機能と作用機序について解析が行われた。得られた成果は以下のとうりである。 (1)本菌が産生する主要なシステインプロテアーゼとして、我々はこれまでの研究からArg-gingipainというユニークな酵素をすでに特定しており、その性状等についても解析を行っているので、本年度の研究では、本酵素の歯周炎における役割とその作用機序に焦点を絞って解析が行われた。その結果、本酵素はオートプロセシングを受けて細胞外へ分泌され、活性型酵素に変換されその病理的機能を発揮することが判った。病原性発現の機序としては、本酵素がタイプIコラーゲンを生理的条件下でよく分解することから、歯周組織の直接破壊に関与することが示唆された。また、本酵素は好中球の活性化を抑え、免疫グロブリンをよく分解することから、生体の防御系の破壊に関与することが示唆された。 (2)本菌が産生するもう一つの主要な菌体外プロテアーゼとして、リジン残基特異的システインプロテアーゼ(Lys-gingipain)を特定した。本酵素の分離精製に成功し、その性状の解析を行った。その結果、本酵素はArg-gingipainとは明らかに異なる性状を有した酵素であるものの、歯周組織の直接破壊や生体防御系の破壊にはArg-gingipainと同様の関与を示すことが明らかとなった。

Research Products
(19 results)

All Other

All Publications (19 results)

[Publications] Tsukuba T.t et al.: "Biochemical properties of the monomeric mutant of human cathepsin E expressed in Chinese hamster ovary cells:Comparison with dimeric forms of the natural and ecombinant cathepsin E." J. Biochem.119. 126-134 (1996)
[Publications] Matsuo K., et al.: "Immunohistochemical localization of cathepsins D and E in human gastric cancer.A possible correlation with local invasive and metastatic activities of carcinoma cells." Human Pathol.27. 184-190 (1996)
[Publications] Kouzuma Y.,et al.: "Purification,characterization,and sequencing of two cysteine proteinase inhibitors,Sca and Scb,from Sunflower(Helianthus annuus)." J.Biochem.119. 1106-1113 (1996)
[Publications] Nakayama K.,et al.: "Involvement of arginie-specific proteinsase(Arg-gingipain)in fimbriation of Porphyromonas gingivalis." J.Bacteriol.178. 2818-2824 (1996)
[Publications] Azuma T.,et al.: "Expression of cathepsin E in pancrease:A possible tumor marker for pancreas,a preliminary report." Int. J. Cancer. 67. 492-497 (1996)
[Publications] Okamoto K.,et al.: "Cloning and sequencing of the gene encoding a novel lysine-specific cysteine proteinase(Lys-gingipain)in Porphyromonas gingivalis:Structural relationship with arginine-specific cysteine proteinase(Arg-gingipain)." J. Biochem.120. 398-406 (1996)
[Publications] Nishishita K.,et al.: "Age-related and dexamethasone-induced changes in cathepsins E and D in rat thymic and splenic cells." Arch.Biochem.Biophys.333. 349-358 (1996)
[Publications] Nakanishi H.,et al.: "Protective effect of MK-801 on the anoxia-aglycemia induced damage in the fluorocitrate-treated hippocampal slice of the rat." Brain Res.732. 232-236 (1996)
[Publications] 山本健二: "プロテアーゼとプロテアーゼインヒビターの新しい機能.創薬研究の新展開を目指して" 日本薬理学会誌. 108. 135-137 (1996)
[Publications] Nakanishi H.,et al.: "Increased expression of cathepsins E and D in neurons of the aged rat barin and their colocalization with lipofuscin and carboxy-terminal fragmnets of Alzheimer amyloid precursor protein." J. Neurochem.68. 739-749 (1997)
[Publications] Kunimats K.,et al.: "Immunohistochemical study of cathepsin G and medullasin in inflamed gingival tissues from periodontal patients." J.Periodont.Res.(in press). (1997)
[Publications] Nakanishi H.,et al.: "Electrophysiological studies of rat substantia nigra neurons in an in vitro slice preparation after middle cerebral artery occulsion." Neuroscience. (in press). (1997)
[Publications] Yamamoto K.,et al.: "Intracellular Protein Catabolism" Plenum Press(New York), 306 (1996)
[Publications] 山本健二他: "Modern Physician" 新興出版社, 24 (1996)
[Publications] Yamamoto K.,et al.: "Medical Aspects of Proteases and Protease Inhibitors" IOS Press(in press), (1997)
[Publications] Yamamoto K.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)
[Publications] Tsukuba T.: "Medical Aspects of Proteases and Protease Inhibitors" IOS Press(in press), (1997)
[Publications] Sakai H.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)
[Publications] Okamoto K.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)

1996 Fiscal Year Annual Research Report

歯周病発症における病原性プロテアーゼの役割と作用機序

Principal Investigator

山本 健二 九州大学, 歯学部, 教授 (40091326)

Research Products

[Publications] Tsukuba T.t et al.: "Biochemical properties of the monomeric mutant of human cathepsin E expressed in Chinese hamster ovary cells:Comparison with dimeric forms of the natural and ecombinant cathepsin E." J. Biochem.119. 126-134 (1996)

[Publications] Matsuo K., et al.: "Immunohistochemical localization of cathepsins D and E in human gastric cancer.A possible correlation with local invasive and metastatic activities of carcinoma cells." Human Pathol.27. 184-190 (1996)

[Publications] Kouzuma Y.,et al.: "Purification,characterization,and sequencing of two cysteine proteinase inhibitors,Sca and Scb,from Sunflower(Helianthus annuus)." J.Biochem.119. 1106-1113 (1996)

[Publications] Nakayama K.,et al.: "Involvement of arginie-specific proteinsase(Arg-gingipain)in fimbriation of Porphyromonas gingivalis." J.Bacteriol.178. 2818-2824 (1996)

[Publications] Azuma T.,et al.: "Expression of cathepsin E in pancrease:A possible tumor marker for pancreas,a preliminary report." Int. J. Cancer. 67. 492-497 (1996)

[Publications] Okamoto K.,et al.: "Cloning and sequencing of the gene encoding a novel lysine-specific cysteine proteinase(Lys-gingipain)in Porphyromonas gingivalis:Structural relationship with arginine-specific cysteine proteinase(Arg-gingipain)." J. Biochem.120. 398-406 (1996)

[Publications] Nishishita K.,et al.: "Age-related and dexamethasone-induced changes in cathepsins E and D in rat thymic and splenic cells." Arch.Biochem.Biophys.333. 349-358 (1996)

[Publications] Nakanishi H.,et al.: "Protective effect of MK-801 on the anoxia-aglycemia induced damage in the fluorocitrate-treated hippocampal slice of the rat." Brain Res.732. 232-236 (1996)

[Publications] 山本健二: "プロテアーゼとプロテアーゼインヒビターの新しい機能.創薬研究の新展開を目指して" 日本薬理学会誌. 108. 135-137 (1996)

[Publications] Nakanishi H.,et al.: "Increased expression of cathepsins E and D in neurons of the aged rat barin and their colocalization with lipofuscin and carboxy-terminal fragmnets of Alzheimer amyloid precursor protein." J. Neurochem.68. 739-749 (1997)

[Publications] Kunimats K.,et al.: "Immunohistochemical study of cathepsin G and medullasin in inflamed gingival tissues from periodontal patients." J.Periodont.Res.(in press). (1997)

[Publications] Nakanishi H.,et al.: "Electrophysiological studies of rat substantia nigra neurons in an in vitro slice preparation after middle cerebral artery occulsion." Neuroscience. (in press). (1997)

[Publications] Yamamoto K.,et al.: "Intracellular Protein Catabolism" Plenum Press(New York), 306 (1996)

[Publications] 山本健二 他: "Modern Physician" 新興出版社, 24 (1996)

[Publications] Yamamoto K.,et al.: "Medical Aspects of Proteases and Protease Inhibitors" IOS Press(in press), (1997)

[Publications] Yamamoto K.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)

[Publications] Tsukuba T.: "Medical Aspects of Proteases and Protease Inhibitors" IOS Press(in press), (1997)

[Publications] Sakai H.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)

[Publications] Okamoto K.,et al.: "Proteolysis and Protein Turnover" IOS Press(in press), (1997)

山本健二九州大学, 歯学部, 教授 (40091326)

[Publications] 山本健二他: "Modern Physician" 新興出版社, 24 (1996)