Antagonistic effect of synthetic peptides corresponding to the binding regions within fimbrial subunit protein from Porphyromonas gingivalis to human gingival fibroblasts.
Specific binding region within fimbrial subunit protein (fimbrilin) from Porphyromonas gingivalis strain 381 was studied in cultured human gingival fibroblasts. Fluorescent micrographs visualised FITC-labelled fimbriae of P. gingivalis specifically bound to normal human fibroblast cell line (Gin-1) along the cell surface. Flow cytometric analysis also revealed the binding of FITC-labelled fimbriae to Gin-1 cells. Synthetic peptides composed of residues 1-20 (AFGVGDDESKVAKLTVMVYN) of the fimbrilin from P. gingivalis, FP381 (1-20), FP381 (69-80; ALTTELTAENQE) and FP381 (171-181; DA-NYLTGSLTT) definitely inhibited P. gingivalis fimbria-binding to Gin-1 cells by enzyme-linked immunosorbent assay (ELISA). Furthermore, based on the Scatchard plot analysis of the binding of 125I-labelled P. gingivalis fimbriae to Gin-1 cells, the apparent dissociation constant (Kd) was calculated as 15.9 pM, and the number of binding sites (Rt) was estimated as 150 sites/cell. Binding studies of 125I-labelled FP381(171-181) also revealed the presence of a non-interacting, single class of affinity binding sites: the apparent Kd and Rt were 29.2 nM and 18440 sites/cell on Gin-1 cells, respectively. These results demonstrate that specific binding regions on P. gingivalis fimbriae to human gingival fibroblasts are present, and certain corresponding peptides clearly inhibited the binding of P. gingivalis fimbriae to human gingival fibroblasts.
Ogawa
T
T
Department of Oral Microbiology, Osaka University Faculty of Dentistry, Japan.
Ogo
H
H
Kinoshita
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000378:metabolism* / Q000494:pharmacology*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000276:immunology; D036781:Fimbriae Proteins; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D005881:Gingiva / Q000166:cytology / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000276:immunology / Q000494:pharmacology*; D018933:Pili, Sex / Q000276:immunology*; D016966:Porphyromonas gingivalis / Q000276:immunology*; D011485:Protein Binding / Q000187:drug effects / Q000276:immunology
D001426:Bacterial Proteins; D010446:Peptide Fragments; C063055:fimbrillin; D036781:Fimbriae Proteins