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Elucidation of molecular mechanism of binding between periodontopathic bacteria and oral streptococci, and development of agents which inhibit forming of dental biofilm

阐明牙周病细菌与口腔链球菌结合的分子机制，开发抑制牙齿生物膜形成的药物

基本信息

批准号：
17390564
负责人：
SHIZUKUISHI Satoshi
金额：
$ 9.98万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390564/
关键词：
periodontopathic bacteria interaction among bacteria biofilm GAPDH fimbriae

项目摘要

In this study, firstly, we examined glyceraldehydes-3-phosphate dehydrogenase (GAPDH) activities on cell surfaces of various oral streptococci which colonize on the tooth surface in the early stage, and their coaggregation activities with Porphyromonas gingivalis. The GAPDH activity on the cell surface of oral streptococci was correlated strongly with the coaggregation activity, therefore, we purified cell surface GAPDHs from five strains of oral streptococci which showed strong cell surface GAPDH and coaggregation activities, and analyzed their interactions with P. gingivalis fimbriae via a biomolecular interaction analysis system (BIAcore). The GAPDHs demonstrated high affinity with P. gingivalis fimbriae and their DNA sequences possessed high degree of homology with one another (more than 95%). Next, we constructed and purified Streptococcus oralis ATCC 9811 recombinant GAPDH (rGAPDH). rGAPDH was digested with various proteinases and applied to reverse-phase HPLC. Peaks were collected and P. gingivalis fimbriae-binding ability of each peak was measured by BIAcore. Amino acid sequence of the fraction which exhibited high binding affinity was determined with mass spectrometer and amino acid analyzer. The binding domain of rGAPDH for P.gingivalis fimbriae existed in amino acid residues 166-183. The synthetic peptide corresponding to the domain inhibited the coaggregation between P.gingivalis and oral streptococci.Moreover, we developed a new method for screening of heterotypic biofilms with a plasmid integration library of Streptococcus gordonii and investigated genes of S.gordonii required to support a heterotypic biofilm community with P.gingivalis. We found S.gordonii governed the development of heterotypic oral biofilms through multiple genetic pathways.

在这项研究中，首先，我们研究了甘油醛-3-磷酸脱氢酶（GAPDH）的活性在各种口腔链球菌的细胞表面的早期阶段，在牙齿表面的殖民，和他们的共聚集活动与牙龈卟啉单胞菌。口腔链球菌的细胞表面GAPDH活性与共聚集活性密切相关，因此，我们从5株具有较强细胞表面GAPDH活性和共聚集活性的口腔链球菌中分离纯化了细胞表面GAPDH，并通过生物分子相互作用分析系统（BIAcore）分析了它们与牙龈卟啉单胞菌菌毛的相互作用。GAPDH与牙龈卟啉单胞菌菌毛具有较高的亲和性，其DNA序列具有较高的同源性（95%以上）。接下来，我们构建并纯化了口腔链球菌ATCC 9811重组GAPDH（rGAPDH）。用各种蛋白酶消化rGAPDH并应用于反相HPLC。收集峰并通过BIAcore测量每个峰的牙龈卟啉单胞菌菌毛结合能力。用质谱仪和氨基酸分析仪对结合亲和力高的组分进行了氨基酸序列测定。rGAPDH与牙龈卟啉单胞菌菌毛的结合域位于第166-183位氨基酸残基。此外，我们建立了一种利用戈登链球菌质粒整合文库筛选异质生物膜的新方法，并对戈登链球菌与牙龈卟啉单胞菌形成异质生物膜群落所需的基因进行了研究。我们发现S.gordonii通过多种遗传途径控制异型口腔生物膜的形成。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

歯周病細菌Porphyromonas gingivalisの口腔内定着における分子メカニズム-初期デンタルバイオフィルム形成菌との相互作用を中心として-,先端歯科医学の創生(浜田茂幸,米田俊之編)

牙周病细菌牙龈卟啉单胞菌口腔定植的分子机制 - 关注与早期牙齿生物膜形成细菌的相互作用 - 高级牙科的创建（由滨田茂之和米田敏之编辑）

DOI：
发表时间：
2005
期刊：
影响因子：
0
作者：
山田聡;村上伸也;Nishiyama Soichiro;Nishiyama Soichiro;Nagata Hideki;Kuboniwa Masae;Guan Su-min;Nagata Hideki;Kuboniwa Masae;Hideki Nagata;永田英樹
通讯作者：
永田英樹

Inhibitory effects of macrocarpals on the biological activity of Porphyromonas gingivalis and other periodontopathic bacteria