PURIFICATION AND ANTIGENICITY OF A NOVEL GLUCAN-BINDING PROTEIN OF STREPTOCOCCUS-MUTANS

A novel glucan-binding protein (GBP) having an apparent molecular mass of 59 kDa (GBP(59)) has been purified from Streptococcus mutans SJ by a combination of affinity chromatography on alpha-1,6-linked glucan, gel filtration chromatography, and ion-exchange chromatography. GBP(59 ) was distinct from the quantitatively predominant S. mutans GBP (GBP( 74)) on the basis of size, elution position in a salt gradient, and an tigenicity. Rat antisera to purified GBP(59) and GBP(74) did not cross -react. GBP(59) is apparently immunogenic in humans, since immunoglobu lin A (IgA) antibody in 20 of 24 adult parotid saliva samples was show n to react with GBP(59) in an enzyme-linked immunosorbent assay. The g lucan-binding activity of GBP(59) was confirmed by anti-GBP(59) immuno gold labelling of Sephadex G-50 that had been preincubated with S. mut ans culture supernatant. GBP(59) could be detected in culture supernat ants of all laboratory strains of S. mutans (e.g., Ingbritt), as well as all strains of S. mutans that had been recently isolated from young children. GBP(59) was often the only component in protease inhibitor- containing 4-h S. mutans culture supernatants that reacted with human parotid salivary IgA antibody in Western blot (immunoblot) analyses. T hese studies suggest that GBP(59) is a structurally and antigenically distinct S. mutans GBP that can elicit significant levels of salivary IgA antibody in humans.