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

共聚体是口腔细菌的定植机制之一。观察到牙龈卟啉单胞菌与中间普雷沃特氏菌共同聚集，不仅在牙周袋中识别，而且在正常牙周缝隙中也能识别。从牙龈假单胞菌的囊泡中分离纯化了一些可能的共聚集因子。从牙龈假单胞菌ATCC 33277的染色体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-HGP17融合蛋白溶液不能聚集中间青霉ATCC 25611。而中间假单胞菌ATCC 25611则与谷胱甘肽Sepharose4B小球结合，后者与GST-Hgp 17蛋白结合。中间假单胞菌ATCC25611分别与GST-HGP44、GST-HGP15、GST-HGP27结合在Glutathione Sepharose4B小球上。HGP44、HGP17、HGP27、HGP15的氨基酸序列具有同源性。中间假单胞菌ATCC 25611与GST-HGP44F(与HGP17和27同源的HGP44内区)和GST-HGP44R(与HGP15同源的HGP44的内区)结合的Glutathiion Sepharose4B小球。抗HgP44的单抗只能微弱地抑制P.的中间产物ATCC 25611与牙龈假单胞菌的囊泡的聚集。

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.