Participation mechanism of periodontopathogenic bacteria into biofilm composed from subgingival bacteria flora with early colonizer

牙周病原菌参与龈下菌群及早期定殖生物膜的机制

• 批准号: 11671808
• 负责人: KAMAGUCHI Arihide
• 金额: $ 2.3万
• 依托单位: Health Sciences University of Hokkaido, School of Dentistry, Department of Oral microbiology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671808/
• 关键词: coaggregation; periodontopathogenic bacteria; adherence; Porphyromonas gingivalis; Prevotella intermedia; rgp A gene; 歯周病原細菌

Coaggregation is one the colonization mechanism of oral bacteria. It is observed that Porphyromonas gingivalis coaggregated with Prevotella intermedia which is recognized not only periodontal pocket but also in normal gingival crevice. Some putative coaggregation factor were purified from the vesicle of P.gingivalis. One of the coaggregation factor gene was cloned from the chromosomal DNA of P.gingivalis ATCC 33277. This putative coaggregation factor gene sequence had homology with HGP 17domain gene of rgp A gene. Fusion protein solution of GST-HGP17protein did not aggregate the P.intermedia ATCC 25611. However, P.intermedia ATCC 25611 adhered to the Glutathione Sepharose 4B bead which bind with GST-HGP 17 protein. P.intermedia ATCC25611 adhere to the Glutathione Sepharose 4B bead which bind with GST-HGP44, GST-HGP15, GST-HGP27, respectively. It had homology of amino acid sequence among HGP44, HGP17, and HGP27 and HGP15. P.intermedia ATCC 25611 adhere to the both Glutathion Sepharose 4B bead which bound GST-HGP44F, internal region of HGP44 which had homology with HGP17 and 27, and GST-HGP44R, internal region of HGP44 which had homology with HGP15. Monoclonal antibody against HGP 44 only weakly inhibited the aggregation of P.intermediate ATCC 25611 with the vesicle of P.gingival is ATCC 33277.These results suggest that outer membrane complex of HGP44, HGP15, HGP17, and HGP27 from rgp A gene of P.gingivalis are associated with the coaggregation with P.intermedia and these complex may have several binding site for P.intermedia.

共聚集是口腔细菌的定植机制之一。牙龈卟啉单胞菌与中间普雷沃菌共聚集，中间普雷沃菌不仅存在于牙周袋中，也存在于正常龈沟中。从牙龈卟啉单胞菌的囊泡中纯化了一些推定的共聚集因子。从牙龈卟啉单胞菌ATCC 33277的染色体DNA中克隆了一个共聚集因子基因。推测的共聚集因子基因序列与rgp A基因的HGP 17结构域基因具有同源性。GST-HGP 17蛋白的融合蛋白溶液不聚集中间毕赤酵母ATCC 25611。然而，中间毕赤酵母ATCC 25611粘附于与GST-HGP 17蛋白结合的Glucose Sepharose 4 B珠。中间毕赤酵母ATCC 25611与Glucose Sepharose 4 B微球结合，分别与GST-HGP 44、GST-HGP 15、GST-HGP 27结合。HGP 44、HGP 17、HGP 27和HGP 15之间氨基酸序列具有同源性。中间青霉ATCC 25611粘附于谷胱甘肽琼脂糖4 B珠，其结合GST-HGP 44 F（与HGP 17和27具有同源性的HGP 44的内部区域）和GST-HGP 44 R（与HGP 15具有同源性的HGP 44的内部区域）。抗HGP 44单克隆抗体仅能微弱地抑制中间型卟啉单胞菌ATCC 25611与牙龈卟啉单胞菌ATCC 33277囊泡的聚集。提示牙龈卟啉单胞菌rgp A基因的HGP 44、HGP 15、HGP 17和HGP 27的外膜复合物与中间型卟啉单胞菌的共聚集有关，这些复合物可能具有多个与中间型卟啉单胞菌结合的位点。