Maeda Kazuhiko, Nagata Hideki, Ojima Miki, Amano Atsuo
Department of Preventive Dentistry, Osaka University Graduate School of Dentistry , Suita, Osaka 565-0871, Japan.
J Proteome Res. 2015 Jan 2;14(1):82-94. doi: 10.1021/pr500848e. Epub 2014 Nov 4.
Porphyromonas gingivalis, a major periodontal pathogen, forms biofilm with other oral bacteria such as streptococci. Here, by using shotgun proteomics, we examined the molecular basis of mixed-biofilm formation by P. gingivalis with Streptococcus oralis. We identified a total of 593 bacterial proteins in the biofilm. Compared to the expression profile in the P. gingivalis monobiofilm, the expression of three proteins was induced and that of 31 proteins was suppressed in the mixed biofilm. Additionally, the expression of two S. oralis proteins was increased, while that of two proteins was decreased in the mixed biofilm, as compared to its monotypic profile. mRNA expression analysis of selected genes using a quantitative reverse transcription polymerase chain reaction confirmed the proteomics data, which included overexpression of P. gingivalis FimA and S. oralis glyceraldehyde-3-phosphate dehydrogenase in association with the biofilm. The results also indicated that S. oralis regulates the transcriptional activity of P. gingivalis luxS to influence autoinducer-2-dependent signaling. These findings suggest that several functional molecules are involved in biofilm formation between P. gingivalis and S. oralis.
牙龈卟啉单胞菌是一种主要的牙周病原体，它会与其他口腔细菌（如链球菌）形成生物膜。在此，我们通过鸟枪法蛋白质组学研究了牙龈卟啉单胞菌与口腔链球菌形成混合生物膜的分子基础。我们在生物膜中总共鉴定出593种细菌蛋白。与牙龈卟啉单胞菌单生物膜中的表达谱相比，混合生物膜中有3种蛋白的表达被诱导，31种蛋白的表达被抑制。此外，与单一型生物膜相比，混合生物膜中两种口腔链球菌蛋白的表达增加，而两种蛋白的表达减少。使用定量逆转录聚合酶链反应对选定基因进行mRNA表达分析，证实了蛋白质组学数据，其中包括牙龈卟啉单胞菌FimA和口腔链球菌甘油醛-3-磷酸脱氢酶与生物膜相关的过表达。结果还表明，口腔链球菌调节牙龈卟啉单胞菌luxS的转录活性，以影响自诱导物-2依赖性信号传导。这些发现表明，几种功能分子参与了牙龈卟啉单胞菌与口腔链球菌之间的生物膜形成。
