Biologically active substances produced by Bacteroides gingivalis and their periodontopathogenicity

牙龈拟杆菌产生的生物活性物质及其牙周致病性

• 批准号: 02404078
• 负责人: NAKAMURA Ryo
• 金额: $ 12.8万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02404078/
• 关键词: Bacteroides gingivalis; Perodontopathogenicity; Hemagglutinin; Trypsin-like enzyme; Band 3 protein; Tissue-cell agglutinin; Cytotoxicity; Vesicle; 赤血球凝集素; B.gingivalis; バクテロイデス・ジンジバリス; 病原因子; 赤血球表面レセプタ-; トリプシン様プロテア-ゼ; 非トリプシン様プロテア-ゼ

Bacteroides gingivalis produces several types of biologically active substances. The purpose of this investigation is to determine the properties of these substances such as proteases, hemagglutinin and tissue-cell agglutinating factor etc. in association with periodontopathogenicity and cytotoxicity.Hemagglutinin (HA), which is considered to participate in the bacterial adherence to host tissues, was isolated and purified from the culture supernatant. The receptor protein of the erythrocyte for this HA was clarified to be band 3 protein and the arginine residue of the peptide was concerned in the binding. This HA possessed both the trypsin-like and the tissue-cell aggregating activities, suggesting that hemagglutination may occur as a result of enzyme-substrate reaction between the trypsin-like enzyme and the band 3 protein.The trypsin-like enzyme produced by B.gingivalis showed the ability of hydrolyzing type I collagen, IgG, IgA and some other naturally occurring proteins. This enzyme also had strong cytotoxicity on human gingival fibroblasts. These facts indicate that the enzyme exhibits pathogenicity not only by destroying periodontal tissue directly but also by decreasing host immune defense ability.All these biologically active substances are concentrated in the secreted outer membrane vesicles of B.gingivalis and all of them behaved in the same way on several protease inhibitors. These results suggest that the true feature of the pathogenic factor of B.gingivalis could be the trypsin-like protease which associates with the outer membrane vesicles.

牙龈拟杆菌产生多种类型的生物活性物质。本研究从牙周组织培养上清中分离纯化了参与细菌粘附的血凝素（Hemagglutinin，HA），并对HA的生物学特性进行了研究。红细胞对该HA的受体蛋白被阐明为带3蛋白，并且肽的精氨酸残基参与结合。该HA具有胰蛋白酶样活性和组织细胞聚集活性，提示其可能通过与带3蛋白的酶-底物反应而发生血凝作用，并具有水解I型胶原、IgG、伊加等天然蛋白的能力。该酶对人牙龈成纤维细胞也有较强的细胞毒性。这些生物活性物质都集中在牙龈B.gingivalis分泌的外膜囊泡中，并且它们对几种蛋白酶抑制剂的作用方式相同。提示牙龈B菌致病因子的真正特征可能是与外膜囊泡结合的胰蛋白酶样蛋白酶。

项目成果

Hayashi,H.: "Survey of a receptor protein in human erythrocytes for hemaggulutinin of Porphyromonas gingivalis." Oral.Microbiol.Immunol.7. 204-211 (1992)

Hayashi,H.：“对人类红细胞中牙龈卟啉单胞菌血凝素受体蛋白的调查。”

森岡昌美: "Porphyromonas gingivalisのヒト歯肉線維芽細胞に対する細胞凝集作用" 口腔衛生学会雑誌. 44. 116-121 (1994)

Masami Morioka：“牙龈卟啉单胞菌对人类牙龈成纤维细胞的细胞聚集作用”口腔健康学会杂志 44. 116-121 (1994)。

森岡 昌美,木戸 玲子,永田 篤司,日野出 大輔,一宮 斉子,佐藤 誠,中村 亮: "Porphyromonas gingivalisのヒト培養歯肉線維芽細胞に対する細胞毒性について" 口腔衛生学会雑誌. 42. 184-190 (1992)

Masami Morioka、Reiko Kido、Atsushi Nagata、Daisuke Hinode、Saiko Ichinomiya、Makoto Sato、Ryo Nakamura：“牙龈卟啉单胞菌对培养的人牙龈成纤维细胞的细胞毒性”口腔健康协会杂志 42. 184-190 (1992)。

Morioka M,Hinode D,Nagata A,Hayashi H,Ichimiya S,Ueda M,Kido R,Nakamura R: "Cytotoxicity of Porphyromonas gingivalis toward tured human gingival fibroblasts" Oral Microbiology and Immunology. 8. (1993)

Morioka M、Hinode D、Nagata A、Hayashi H、Ichimiya S、Ueda M、Kido R、Nakamura R：“牙龈卟啉单胞菌对人牙龈成纤维细胞的细胞毒性”口腔微生物学和免疫学。

日野出 大輔,林 祐行,嶋田 順子,萬好都 美恵,中村 亮: "Porphyromonas gingivalisの産生する非トリプシン様酵素の性状。" 口腔衛生学会雑誌. 41. 478-479 (1991)

Daisuke Hinode、Yuyuki Hayashi、Junko Shimada、Mie Mankoto、Ryo Nakamura：“牙龈卟啉单胞菌产生的非胰蛋白酶样酶的特性。”口腔卫生学会杂志 41. 478-479 (1991)。