Development of inhibitory peptide in oral bacterial coaggregation

本研究的目的是确定介导牙龈嗜水单胞菌与口腔革兰氏阳性菌共聚集的共粘附素，并在共聚集过程中产生抑制肽。浊度法测定，在牙龈假单胞菌的各组分中，菌毛对牙龈假单胞菌与口腔链球菌的共聚集有较强的抑制作用。重组纤布蛋白和合成肽的抑制研究表明，纤布蛋白的氨基酸残基266 ~ 286包含一个参与共聚集的结构域。在此之前，我们报道了该结构域介导牙龈卟啉卟啉与富含脯氨酸蛋白（praline-rich protein， REP）和淀粉蛋白的结合。因此，该肽可能是一种有效的抗牙龈假单胞菌粘附的抑制剂。我们也报道了PRP的羧基末端片段负责rep与牙龈卟啉的结合。在这项研究中，我们发现该片段抑制了牙龈假单胞菌对形成斑块的革兰氏阳性细菌的粘附，以及对PRP的粘附。该片段似乎也是一种有效的聚集抑制肽。为了鉴定革兰氏阳性菌的共黏附素，研究了口腔链球菌表面成分与牙龈假单胞菌膜的相互作用。35 kda口腔表面组分与牙龈假单胞菌菌毛发生反应。部分纯化的35-kDa蛋白抑制牙龈假单胞菌和口腔假单胞菌的共聚集。35-kDa蛋白抗体与血链球菌和哥氏链球菌反应，表明多种口腔革兰氏阳性菌中均存在与35-kDa表面蛋白相关的类似蛋白。它们似乎在共聚集中也起着重要作用。进一步鉴定牙龈假单胞菌在35 kda蛋白中的结合区正在进行中。在这项研究中，我们获得了几种抑制牙龈菌对革兰氏阳性细菌粘附的肽。虽然这些肽的进一步评价是必要的，他们可能是可用的，以防止牙龈粘附牙菌斑。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