Development of inhibitory peptide in oral bacterial coaggregation

本研究的目的是确定介导牙龈假单胞菌与口腔革兰氏阳性菌共聚集的粘附素，并在共聚集中开发抑制肽。用比浊法测定，在牙龈假单胞菌的各种成分中，菌毛对牙龈假单胞菌和口腔链球菌的共聚集有很强的抑制作用。利用重组芬布林和合成肽进行的抑制研究表明，菲莫林的266-286位氨基酸残基含有一个参与共聚集的结构域。在此之前，我们曾报道过该结构域介导牙龈假单胞菌与富含脯氨酸的蛋白(Rep)和他汀类药物的结合。因此，该多肽可能是一种有效的抑制牙龈假单胞菌黏附的药物。我们还报道了PRP的羧基末端片段负责与牙龈假单胞菌的rep结合。在这项研究中，我们发现该片段抑制了牙龈假单胞菌对菌斑形成的革兰氏阳性细菌和…的黏附更重要的是，这个片段似乎也是一种有效的共聚集抑制肽。为了从革兰氏阳性菌中鉴定粘附素，研究了口腔葡萄球菌表面成分与牙龈假单胞菌表面成分的相互作用。35 kDa口腔表面成分与牙龈假单胞菌菌毛发生反应。部分纯化的35 kDa蛋白抑制了牙龈假单胞菌与金黄色葡萄球菌之间的共聚集。抗35-kDa蛋白抗体与血链球菌和戈登链球菌反应，表明口腔革兰氏阳性菌中存在与35-kDa口腔葡萄球菌表面蛋白相似的蛋白。它们似乎也在共同聚集中发挥了重要作用。进一步鉴定35-kDa蛋白中的牙龈假单胞菌结合区正在进行中。在本研究中，我们获得了几种抑制牙龈假单胞菌与革兰氏阳性菌黏附的多肽。尽管有必要对这些多肽进行进一步的评估，但它们可能有助于防止牙龈假单胞菌附着在牙菌斑上。较少

The objects of this study are to determine the coadhesins which mediate coaggregation between Pomhyromonas gingivalis and oral gram-positive bacteria, and to develop inhibitory peptide in the coaggregation. Among various components of P.gingivalis, fimbriae strongly inhibited the coaggregation between P.gingivalis and Streptococcus oralis by the turbidimetric method. The inhibition studies using recombinant fimbrillin and synthetic peptides showed that amino acid residues 266-286 of fimbrillin contain a domain involved in the coaggregation. Previously, we reported that this domain mediates the binding of P.gingivalis to praline-rich protein (REP) and statherin. Therefore, this peptide may acy as an effective inhibitor against P.gingivalis adherence. We also reported that the carboxyl-terminal fragment of PRP is responsible for the REP-binding to P.gingivalis. In this study, we found that this fragment inhibited P.gingivalis adherence to plaque-forming gram-positive bacteria as well as … More to PRP.This fragment also appears to be one of the potent coaggregation-inhibitory peptides. To identify the coadhesin from gram-positive bacteria, the interactions of surface components from S.oralis with P.gingivalis flmbriae were examined. 35-kDa S.orals surface component reacted with P.gingivalis fimbriae. The partial purified 35-kDa protein inhibited the coaggregation between P.gingivalis and S.oralls. The antibodies against 35-kDa protein reacted with Streptococcus sanguis and Streptococcus gordonii, indicating that the similar proteins related to 35-kDa S.oralis surface protein exsit in various oral gram-positive bacteria. They also seems to play an important role in the coaggregation. Further identification of the P.gingivalis binding region in the 35-kDa protein is in progress. In-this study, we obtained several peptides which inhibit P.gingivalls adherence to gram-positive bacteria. Although further evaluation of these peptides are necessary, they may be avairable to prevent P.gingivalls adherence to dental plaque. Less