Identification of glyceraldehyde-3-phosphate dehydrogenase of epithelial cells as a second molecule that binds to Porphyromonas gingivalis fimbriae.
Binding of Porphyromonas gingivalis to the host cells is an essential step in the pathogenesis of periodontal disease. P. gingivalis binds to and invades epithelial cells, and fimbriae are thought to be involved in this process. In our earlier studies, two major epithelial cell components of 40 and 50 kDa were identified as potential fimbrial receptors. Sequencing of a cyanogen bromide digestion fragment of the 50-kDa component resulted in an internal sequence identical to keratin I molecules, and hence this cytokeratin represents one of the epithelial cell receptors for P. gingivalis fimbriae. In this study, the 40-kDa component of KB cells was isolated and its amino-terminal sequence determined. The N-terminal amino sequence was found to be GKVKVGVNGF and showed perfect homology with human glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Furthermore, purified P. gingivalis fimbriae were found to bind to rabbit muscle GAPDH. Antibodies directed against internal peptide 49-68 and 69-90 of fimbrillin were shown to inhibit the binding of P. gingivalis and of fimbriae to epithelial cells. Antibodies against these peptides also inhibited the binding of fimbriae to GAPDH. Our results confirmed that the amino-terminal domain corresponding to amino residues 49-68 of the fimbrillin protein is the major GAPDH binding domain. These studies point to GAPDH as a major receptor for P. gingivalis major fimbriae and, as such, GAPDH likely plays a role in P. gingivalis adherence and colonization of the oral cavity, as well as triggering host cell processes involved in the pathogenesis of P. gingivalis infections.
Sojar
Hakimuddin T
HT
Department of Oral Biology, School of Dental Medicine, State University of New York at Buffalo, 213 Foster Hall, 3435 Main Street, Buffalo, NY 14214-3092, USA. orbhakim@buffalo.edu
Genco
Robert J
RJ
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial; D001422:Bacterial Adhesion; D004847:Epithelial Cells / Q000201:enzymology*; D036781:Fimbriae Proteins / Q000276:immunology; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007624:KB Cells; D008969:Molecular Sequence Data; D016966:Porphyromonas gingivalis / Q000472:pathogenicity / Q000502:physiology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
D000907:Antibodies, Bacterial; C063055:fimbrillin; D036781:Fimbriae Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases