Proteolysis of the 185,000 MW streptococcal cell wall antigen generating 4000 and 6000 MW peptides with distinct antigenic determinants.

A 185,000 MW glycoprotein antigen derived from Streptococcus mutans was digested with subtilisin. Purification by reversed phase high-powered liquid chromatography (HPLC) resulted in a homogeneous 20,000 MW protein which possesses the streptococcal antigen (SA) I and II determinants. This protein was immunogenic in mice both for the 20,000 MW and the native 185,000 MW SA. Further proteolysis with subtilisin generated four peptides of 18,000, 10,000, 6000 and 4000 MW. Whereas the 20,000, 18,000 and 10,000 MW peptides contained both SAI and SAII determinants, the 4000 MW peptide possessed only the SAI and the 6000 MW peptide the SAII determinant. The 4000-SAI peptide is of special significance, as the smallest SAI material separated in the past was 150,000 MW. This was difficult to purify and proved to be protective against dental caries on immunization of rhesus monkeys.