IDENTIFICATION OF LINEAR ANTIGENIC SITES ON THE PORPHYROMONAS-GINGIVALIS 43-KDA FIMBRILLIN SUBUNIT

The fimbrillin of Porphyromonas gingivalis is thought to be an importa nt virulence factor that mediates adherence to host surfaces. The line ar immunogenic and antigenic structure of P. gingivalis fimbrillin was investigated with synthetic peptides corresponding to the amino acid sequence predicted from the cloned fimbrillin gene for P. gingivalis 2 561. A series of continuous and overlapping peptides corresponding to the entire sequence of P. gingivalis fimbrillin was used to immunize W istar rats. The resulting polyclonal antibodies were used to test the antigenicity of the 43-kDa fimbrillin protein by enzyme-linked immunos orbent assay and Western blot analysis. All the peptides elicited spec ific antibodies directed to the corresponding peptides but differed in their ability to elicit antisera that cross-reacted with either nativ e or denatured fimbrillin. Antisera to various C-terminal one-third pe ptides were more reactive to the denatured monomeric form of fimbrilli n by Western blot analysis. Antisera to peptide 99-110 was by far the most reactive against the native form of the oligomeric fimbrillin as well as the partially denatured oligomeric form of fimbrillin. The res ults indicate that amino acid residues 99-110 on the native fimbrillin protein are accessible to antibody binding and that the immunogen 99- 110, when conjugated to thyroglobulin, is able to mimic an epitope on the 43-kDa fimbrillin.