Effect of Acidic pH on Expression of Surface-Associated Proteins of Streptococcus oralis

Abstract

Streptococcus oralis, a member of the mitis group of oral streptococci, is implicated in the pathogenesis of infective endocarditis and is the predominant aciduric non-mutans-group streptococcus in dental plaque. We undertook to identify the most abundant surface-associated proteins of S. oralis and to investigate changes in protein expression when the organism was grown under acidic culture conditions. Surface-associated proteins were extracted from cells grown in batch culture, separated by two-dimensional gel electrophoresis, excised, digested with trypsin, and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry and liquid chromatography-tandem mass spectrometry. Putative functions were assigned by homology to a translated genomic database of Streptococcus pneumoniae. A total of 27 proteins were identified; these included a lipoprotein, a ribosome recycling factor, and the glycolytic enzymes phosphoglycerate kinase, fructose bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, and enolase. The most abundant protein, phosphocarrier protein HPr, was present as three isoforms. Neither lactate dehydrogenase nor pyruvate oxidase, dominant intracellular proteins, were present among the proteins on the gels, demonstrating that proteins in the surface-associated pool did not arise as a result of cell lysis. Eleven of the proteins identified were differentially expressed when cells were grown at pH 5.2 versus pH 7.0, and these included superoxide dismutase, a homologue of dipeptidase V from Lactococcus lactis, and the protein translation elongation factors G, Tu, and Ts. This study has extended the range of streptococcal proteins known to be expressed at the cell surface. Further investigations are required to ascertain their functions at this extracellular location and determine how their expression is influenced by other environmental conditions.Streptococcus oralis, a member of the mitis group of the viridans group streptococci, is a component of normal dental plaque, in which it forms a significant proportion of the aciduric microflora (5, 34). Aciduricity, the ability to grow under conditions of low pH, is considered a virulence determinant for bacteria associated with the initiation and progression of dental caries, although the precise role of S. oralis in this disease process has yet to be fully defined. In addition to the potential role of S. oralis in dental caries, it is well documented that S. oralis is associated with a range of extraoral diseases including endocarditis and infections in susceptible patients, including those who are immunologically compromised. Douglas and coworkers (13) documented that, out of 42 cases of infective endocarditis from which viridans group streptococci were isolated, the most common species were from the mitis group, namely, Streptococcus sanguis, S. oralis, and Streptococcus gordonii (isolation frequencies of 31.9, 29.8, and 12.7%, respectively). S. oralis is also the predominant species of the viridans group streptococci as...

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