Analysis of major outer membrane proteins from periodontopathogen and relationship between these proteins and clinical symptom in periodontitis

牙周病原菌主要外膜蛋白分析及其与牙周炎临床症状的关系

• 批准号: 15591958
• 负责人: MURAKAMI Yukitaka
• 金额: $ 2.18万
• 依托单位: Aichi-Gakuin University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591958/
• 关键词: periodontopathogen; Porphyromonas gingivalis; major outer membrane protein; environmental factors; continuous culture; 2-dimensional gel electrophoresis; deletion mutants; sera from periodontitis patients; 培養条件; 培養温度; OmpA; 2次元電気泳動法; RagA; Ra

1.Analysis of major outer membrane proteins among various P.gingivalis strains.Major outer membrane proteins, such as RagA and Pgm6/7,were found in all strains tested in a similar distribution pattern. However, mobility and reaction of RagB against specific antibody was different among strains, indicating RagB heterogeneity.2.Effect of culture environment factors on the expression of P.gingivalis major outer membrane proteinsWhen effect of several factors was examined on the expression of major outer membrane proteins, temperature was the most important factor. As temperature increased, expression of RagA,RagB and gingipains as well as fimbriae and short fimbriae decreased.Then, we used continuous culture in which regulation of culture environment was possible. With increase of growth rate, production of fimbriae and short fimbriae increased, while amount and activity of gingipains decreased. In a nutrient-poor medium, fimbriae and short fimbriae decreased. When atmosphere was changed … More from anaerobic to aerobic condition, activity of gingipains decreased. Protein spots whose amount increased under oxidative stress was observed by 2-dimensional gel electrophoresis, and those were identified by mass spectrometry3.Characterization of major outer membrane proteins using gene-deletion mutants.Major outer membrane proteins Pgm6/7, which were homologous to OmpA protein from Escherichia coli, existed in outer membrane forming heterotrimers. Since numerous vesicle-like structures were observed in the case of Pgm6/7 gene-deletion mutants by electron microscopy, Pgm6/7 might be involved in stabilization of outer membrane integrity. Since construction of RagA and RagB-deletion mutants have been finished, we are characterizing the mutants.4.Immunological reactions of sera from patients with periodontitis to P.gingivalis major outer membrane proteins.To examine immunoreactive antigens in P.gingivalis, sera from patients with periapical periodontitis were used. More than half of the sera showed strong immunological reaction to P.gingivalis cell components, especially, outer membrane proteins RagB and gingipains. Clinical symptoms such as percussion and periapical translucency were related to immnoreactivity to P.gingivalis. Less

1.不同牙龈卟啉单胞菌菌株主要外膜蛋白的分析在所有测试的菌株中发现主要外膜蛋白，如RagA和Pgm 6/7，其分布模式相似。但不同菌株间RagB的迁移率和对特异性抗体的反应性不同，表明RagB具有异质性。2.培养环境因素对牙龈卟啉单胞菌主要外膜蛋白表达的影响当考察多种因素对主要外膜蛋白表达的影响时，温度是最重要的因素。随着温度的升高，RagA、RagB和牙龈卟啉酶的表达以及菌毛和短菌毛的表达均下降。随着生长速度的增加，菌毛和短菌毛的产量增加，牙龈卟啉酶的数量和活性下降。在营养不良的培养基中，菌毛和短菌毛减少。当气氛改变时 ...更多信息 从厌氧到有氧，牙龈卟啉菌蛋白酶活性下降。通过双向凝胶电泳观察氧化应激条件下蛋白质点数量增加的情况，并通过质谱鉴定。3.利用基因缺失突变体表征主要外膜蛋白质Pgm 6/7，与大肠杆菌OmpA蛋白同源，存在于外膜中，形成异源三聚体。由于许多囊泡样结构的情况下，通过电子显微镜观察到Pgm 6/7基因缺失突变体，Pgm 6/7可能参与外膜完整性的稳定。由于RagA和RagB缺失突变体的构建已经完成，我们正在对突变体进行表征。4.牙周炎患者血清对牙龈卟啉单胞菌主要外膜蛋白的免疫反应。半数以上的血清对牙龈卟啉单胞菌的细胞成分，尤其是外膜蛋白RagB和牙龈卟啉单胞菌蛋白酶有较强的免疫反应。临床症状，如叩诊和根尖周炎与牙龈卟啉单胞菌的免疫反应性。少

项目成果

A comparison of gingipains production by the periodontal pathogenic bacterium Porphyromonas gingivalis at different growth rates in a continuous flow culture system (in Japanese)

连续流动培养系统中牙周病原菌牙龈卟啉单胞菌在不同生长速率下产生牙龈蛋白酶的比较（日语）

• 发表时间: 2003
• 期刊: Journal of Japanese Association for Anaerobic Infection Research Vol.33
• 作者: Masuda;T. et al.
• 通讯作者: T. et al.

The short fimbriae of Porphyrmonas gingivalis and their role in coadhesion with Streptococcus gordonii

牙龈卟啉单胞菌的短菌毛及其在与戈登链球菌粘附中的作用

• 发表时间: 2005
• 期刊: Infect.Immun. 73・7
• 作者: Park;Y.et al.
• 通讯作者: Y.et al.

Major outer membrane proteins from Porphyromonas gingivalis:: strain variation, distribution, and clinical significance in periradicular lesions

• DOI: 10.1111/j.1600-0722.2005.00235.x
• 发表时间: 2005-10-01
• 期刊: EUROPEAN JOURNAL OF ORAL SCIENCES
• 影响因子: 1.9
• 作者: Imai, M;Murakami, Y;Yoshimura, F
• 通讯作者: Yoshimura, F

The trimeric structure of major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis

牙龈卟啉单胞菌中与OmpA同源的主要外膜蛋白的三聚体结构

• 发表时间: 2005
• 期刊: J.Bacteriol. 187・3
• 作者: Nagano;K. et al.
• 通讯作者: K. et al.

Analysis of major virulence factors in Porphyromonas gingivalis under various culture temperature using specific antibodies

特异性抗体分析不同培养温度下牙龈卟啉单胞菌主要毒力因子

• 发表时间: 2004
• 期刊: Microbiol.Immunol. Vol.48
• 作者: Murakami;Y. et al.
• 通讯作者: Y. et al.