Ray C A, Gfell L E, Buller T L, Gregory R L
Department of Oral Biology, School of Dentistry, Indiana University, Indianapolis, Indiana 46202-5186, USA.
Clin Diagn Lab Immunol. 1999 May;6(3):400-4. doi: 10.1128/CDLI.6.3.400-404.1999.
Streptococcus mutans has been implicated as the major causative agent of human dental caries. S. mutans binds to saliva-coated tooth surfaces, and previous studies suggested that fimbriae may play a role in the initial bacterial adherence to salivary components. The objectives of this study were to establish the ability of an S. mutans fimbria preparation to bind to saliva-coated surfaces and determine the specific salivary components that facilitate binding with fimbriae. Enzyme-linked immunosorbent assay (ELISA) established that the S. mutans fimbria preparation bound to components of whole saliva. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot techniques were used to separate components of whole saliva and determine fimbria binding. SDS-PAGE separated 15 major protein bands from saliva samples, and Western blot analysis indicated significant binding of the S. mutans fimbria preparation to a 52-kDa salivary protein. The major fimbria-binding salivary protein was isolated by preparative electrophoresis. The ability of the S. mutans fimbria preparation to bind to the purified salivary protein was confirmed by Western blot analysis and ELISA. Incubation of the purified salivary protein with the S. mutans fimbria preparation significantly neutralized binding of the salivary protein-fimbria complex to saliva-coated surfaces. The salivary protein, whole saliva, and commercial amylase reacted similarly with antiamylase antibody in immunoblots. A purified 65-kDa fimbrial protein was demonstrated to bind to both saliva and amylase. These data indicated that the S. mutans fimbria preparation and a purified fimbrial protein bound to whole-saliva-coated surfaces and that amylase is the major salivary component involved in the binding.
变形链球菌被认为是人类龋齿的主要致病因素。变形链球菌可黏附于唾液包被的牙齿表面,先前的研究表明菌毛可能在细菌对唾液成分的初始黏附中起作用。本研究的目的是确定变形链球菌菌毛制剂与唾液包被表面的结合能力,并确定促进与菌毛结合的特定唾液成分。酶联免疫吸附测定(ELISA)表明,变形链球菌菌毛制剂可与全唾液成分结合。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹技术分离全唾液成分并确定菌毛结合情况。SDS-PAGE从唾液样本中分离出15条主要蛋白带,蛋白质印迹分析表明变形链球菌菌毛制剂与一种52 kDa的唾液蛋白有显著结合。通过制备电泳分离出主要的菌毛结合唾液蛋白。蛋白质印迹分析和ELISA证实了变形链球菌菌毛制剂与纯化唾液蛋白的结合能力。纯化的唾液蛋白与变形链球菌菌毛制剂孵育可显著中和唾液蛋白-菌毛复合物与唾液包被表面的结合。在免疫印迹中,唾液蛋白、全唾液和商业淀粉酶与抗淀粉酶抗体的反应相似。一种纯化的65 kDa菌毛蛋白被证明可与唾液和淀粉酶结合。这些数据表明,变形链球菌菌毛制剂和一种纯化的菌毛蛋白可与全唾液包被的表面结合,且淀粉酶是参与结合的主要唾液成分。