High resolution structure analysis of Porphyromonas gingivalis membrane proteins in the biofilm.

生物膜中牙龈卟啉单胞菌膜蛋白的高分辨率结构分析。

• 批准号: 22792114
• 负责人: MAEDA Kazuhiko
• 金额: $ 2.58万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22792114/
• 关键词: 予防歯科学; 歯周病; Porphyromonas gingivalis; 口腔レンサ球菌; プロテオーム; バイオフィルム; Prophyromonas gingivalis; 膜タンパク質; GAPDH

The purpose of this study is to identify and characterize new Porphyromonas gingivalis components that interact with Streptococcus oralis GAPDH. Pull-down assay was performed to detect potential interactions between P. gingivalis client proteins and S. oralis recombinant GAPDH. The proteins were separated by 2D-gel electrophoresis, and the five proteins ; RagA4, AbfD, GAPDH, GDH and MDH were identified by a proteomic-tandem mass spectrometry. Interaction between these P. gingivalis client proteins and S. oralis GAPDH was analyzed via a biomolecular interaction analysis system. S. oralis GAPDH demonstrated high affinity with five client proteins. Coaggregation between P. gingivalis and S. oralis was measured by turbidimetric method and fluorescence microscope. Recombinant RagA4, AbfD and GDH enhanced the coaggregation, whereas recombinant GAPDH and MDH inhibited in a dose-dependent manner. Furthermore, recombinant RagA4, AbfD and GDH regulated expression of luxS in P. gingivalis. These results indicate that five client membrane proteins of P. gingivalis might be regulators in P. gingivalis biofilm formation with oral streptococci.

本研究的目的是鉴定和表征与口腔链球菌GAPDH相互作用的新牙龈卟啉单胞菌组分。Pull-down检测牙龈卟啉单胞菌客户蛋白和沙门氏菌之间的潜在相互作用。口腔腺病毒重组GAPDH。通过2D-凝胶电泳分离蛋白质，并通过蛋白质组学-串联质谱鉴定了5种蛋白质：RagA 4，AbfD，GAPDH，GDH和MDH。这些牙龈卟啉单胞菌客户蛋白与S.通过生物分子相互作用分析系统分析口腔GAPDH。S.口腔菌GAPDH与5个客户蛋白具有高亲和力。牙龈卟啉单胞菌与S.采用比浊法和荧光显微镜对口腔粘膜进行定量测定。重组RagA 4，AbfD和GDH增强共聚集，而重组GAPDH和MDH以剂量依赖性方式抑制。此外，重组RagA 4、AbfD和GDH在牙龈卟啉单胞菌中调节luxS的表达。这些结果表明，牙龈卟啉单胞菌的五个客户膜蛋白可能是牙龈卟啉单胞菌与口腔链球菌形成生物膜的调节剂。

项目成果

Porphyromonas gingivalis NAD-dependent glutamate dehydrogenase binds to glyceraldehyde-3-phosphate dehydrogenase of Streptococcus oralis

牙龈卟啉单胞菌 NAD 依赖性谷氨酸脱氢酶与口腔链球菌的甘油醛-3-磷酸脱氢酶结合

• 发表时间: 2011
• 作者: Maeda Kazuhiko;Nagata Hideki;Kuboniwa Masae;Ojima Miki
• 通讯作者: Ojima Miki