Department of Preventive Dentistry, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, 770-8504, Japan.
Infect Immun. 2010 Mar;78(3):1185-92. doi: 10.1128/IAI.01224-09. Epub 2009 Dec 14.
We have previously shown that one of the minimal active regions of statherin, a human salivary protein, for binding to Fusobacterium nucleatum is a YQPVPE amino acid sequence. In this study, we identified the FomA protein of F. nucleatum, which is responsible for binding to the statherin-derived YQPVPE peptide. Overlay analysis showed that a 40-kDa protein of the F. nucleatum cell envelope (40-kDa CE) specifically bound to the YQPVPE peptide. The equilibrium association constant between the affinity-purified 40-kDa CE and the YQPVPE peptide was 4.30 x 10(6). Further, the purity and amino acid sequence analyses of the purified 40-kDa CE revealed approximately 98.7% (wt/wt) purity and a high degree of homology with FomA, a major porin protein of F. nucleatum. Thus, a FomA-deficient mutant failed to bind to the YQPVPE peptide. In addition, increased levels of a FomA-specific mucosal IgA antibody (Ab) and plasma IgG and IgA Abs were seen only in mice immunized nasally with cholera toxin (CT) and the purified 40-kDa FomA protein. Interestingly, saliva from mice that received FomA plus CT as a mucosal adjuvant nasally prevented in vitro binding of F. nucleatum to statherin-coated polyvinyl chloride plates. Taken together, these results suggest that induction of specific immunity to the 40-kDa FomA protein of F. nucleatum, which specifically binds to the statherin-derived peptide, may be an effective tool for preventing the formation of F. nucleatum biofilms in the oral cavity.
我们之前已经表明,唾液蛋白牙固蛋白的一个最小活性区域是一个与 Fusobacterium nucleatum 结合的 YQPVPE 氨基酸序列。在这项研究中,我们鉴定了 F. nucleatum 的 FomA 蛋白,它负责与牙固蛋白衍生的 YQPVPE 肽结合。覆盖分析显示,F. nucleatum 细胞包膜的 40-kDa 蛋白(40-kDa CE)特异性结合到 YQPVPE 肽上。亲和纯化的 40-kDa CE 与 YQPVPE 肽之间的平衡缔合常数为 4.30 x 10(6)。此外,纯化的 40-kDa CE 的纯度和氨基酸序列分析表明,其纯度约为 98.7%(wt/wt),并且与 F. nucleatum 的主要孔蛋白 FomA 具有高度同源性。因此,FomA 缺陷型突变体不能与 YQPVPE 肽结合。此外,仅在鼻腔用霍乱毒素(CT)和纯化的 40-kDa FomA 蛋白免疫的小鼠中观察到 FomA 特异性粘膜 IgA 抗体(Ab)和血浆 IgG 和 IgA Ab 的水平增加。有趣的是,鼻腔接受 FomA 加 CT 作为粘膜佐剂的小鼠的唾液防止了 Fusobacterium nucleatum 在涂有牙固蛋白的聚氯乙烯板上的体外结合。总之,这些结果表明,诱导对 F. nucleatum 的 40-kDa FomA 蛋白的特异性免疫,该蛋白特异性结合牙固蛋白衍生的肽,可能是预防口腔中 Fusobacterium nucleatum 生物膜形成的有效工具。