Development studies on specific inhibitors of adherence of periodontal pathogen based on the etiology

牙龈卟啉单胞菌是一种革兰氏阴性厌氧菌，与牙周炎的发病有关。牙龈卟啉单胞菌伞毛被认为在牙周组织的定植和侵袭中发挥重要作用。我们分析了牙龈卟啉单胞菌菌毛与人细胞外基质蛋白（即，例如，纤连蛋白、层粘连蛋白、胶原蛋白），血红蛋白，纤维蛋白原和唾液成分（即，富含脯氨酸的蛋白质[PRP]、富含脯氨酸的糖蛋白[PRG]和雌甾苷肽）。观察到菌毛与我们使用的所有人蛋白质具有高亲和力。来自牙龈卟啉单胞菌的精氨酸特异性蛋白酶可以暴露基质蛋白中的C-末端Arg残基，并导致该生物体的增强的菌毛介导的结合。我们用亲和柱层析法检测了菌毛与一些合成二肽的结合。仅观察到Gly-Arg配体 关于我们 与菌毛有亲和性。此外，我们分析了菌毛和固定化纤连蛋白（完整或部分降解的纤连蛋白的蛋白酶）的相互作用。BlAcore图谱表明菌毛和蛋白酶降解的纤连蛋白之间的相互作用增强。这种增强作用被L-Arg或包括缓激肽在内的C-末端含有Arg残基的寡肽所抑制，表明牙龈卟啉单胞菌菌毛可能具有与C-末端Arg残基紧密结合的能力。PRP-C肽是一种有效的PRP-菌毛相互作用抑制剂，它对应于PRP的C-末端由21个氨基酸残基组成的片段，显著抑制菌毛与唾液蛋白和胶原的结合，并且对BlAcore上的纤连蛋白和层粘连蛋白的抑制作用较小。这些观察结果表明，P. givalis菌毛通过不同的机制与宿主蛋白质相互作用，最终促进细菌在口腔中的粘附。少

Porphyromonas gingivalis, a Gram-negative anaerobe, is known to be involved in the pathogenesis of periodontitis. P.gingivalis fimbriae are thought to play an important role in the colonization and invasion to periodontal tissues. We analyzed the interactions of P.gingivalis fimbriae with human extracellar matrix proteins (i. e., fibronectin, laminin, collagen), hemoglobin, fibrinogen and salivary components (i.e., proline-rich protein [PRP], proline-rich glycoprotein [PRG] and statherin) based on surface plasmon resonance (SPR) spectroscopy using a biomolecular interaction analyzing system (BlAcore). Fimbriae were observed to have high affinity with all the human proteins we used. Arginine-specific protease from P.gingivalis can expose the C-terminal Arg residue in the matrix proteins, and results in enhanced fimbria-mediated binding of this organism. We examined the binding of fimbriae to some synthetic dipeptides by using affinity column chromatography. Only Gly-Arg ligand was obser … More ved to have affinity with fimbriae. In addition, we analyzed the interaction of fimbriae and immobilized fibronectins (intact or partially degraded fibronectin by the protease). BlAcore profiles demonstrated an enhanced interaction between fimbriae and protease-degraded fibronectin. This enhancement was inhibited by the addition of L-Arg, or oligopeptides including bradykinin containing the Arg residue at the C-terminus, suggesting that the P.gingivalis fimbriae may potentially have an ability to bind tightly to the C-terminal Arg residue. The peptide PRP-C, which is corresponding to the C-terminal segment composed of 21 amino acid residues of PRP, and a potent inhibitor of PRP-fimbriae interaction, dramatically inhibited the fimbrial association to the salivery proteins and collagen, and was less inhibitory against fibronectin and laminin on BlAcore. These observations suggested that P.gin givalis fimbriae interact diversly with the host proteins by different mechanisms, which eventually promotes the bacterial adherence to the oral cavity. Less