UBC Theses and Dissertations
Cell surface of hydrophobic and hydrophilic strains of Streptococcus sanguis Ganeshkumar, Nadarajah
Cell surfaces of aggregation, adherence or hydrophilic variants of Streptococcus sanguis were compared to cell surfaces of the parent strain with regard to their protein and antigenic constituents. Cell surface molecules were released by digestion with mutanolysin. Extraction with SDS-BME, urea, lithium chloride and boiling water did not solubilize any material which stained with silver nitrate in an SDS-polyacrylamide gel. The parent organism S. sanguis 12 which aggregates in saliva, adheres to saliva coated hydroxyapatite (S-HA) and is hydrophobic was found to possess a prominently staining 160,000 MW protein. This protein was almost completely absent from strain I2na, a hydrophobic non-aggregating variant, and was completely absent from the hydrophilic non-aggregating, non-adherent strain 12L. Trypsinization of strain 12 resulted in the coincident loss of the 160,000 MW protein and the ability to aggregate in saliva. Trypsin treatment reduced, but did not eliminate the hydrophobic character of the cells. Boiling destroyed the ability to aggregate but did not alter hydrophobicity. Cell wall digests of strain 12 contained a large number of proteins which were absent from strains 12na and 12L. Mutanolysin digests of the hydrophilic strains contained no material that was visible in a silver stained SDS-polyacrylamide gel. Electron microscopy of phosphotungstic acid stained cells showed a thick capsular material spread evenly over the cell surface of the parent strain 12. This layer was thinner around the cells of strain 12na and appeared patchy on hydrophilic strains. Electron microscopy of uranyl acetate stained cells revealed that strain 12 had short fibrillar structures evenly distributed over the cell surface, and long fibrils which were more concentrated at the end of the cell. The hydrophilic strain 12L lacked both types of fibrils. Crossed immunoelectrophoresis confirmed that the major cell surface antigens were located in the 160,000 MW region. Cell wall digests of strain 12 and 12na inhibited adherence of strain 12 to S-HA by 36% and 19% respectively. The digests of hydrophilic strain 12L were not inhibitory. The inhibitory activity was sensitive to heat and SDS.
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