Selection and phenotypic characterization of nonhemagglutinating mutants of Porphyromonas gingivalis.
To further investigate the relationship between fimbriae and the hemagglutinating adhesin HA-Ag2 of Porphyromonas gingivalis, three spontaneous mutants of the type strain ATCC 33277 were selected by a hemadsorption procedure. They were characterized for hemagglutination, trypsin-like and lectin-binding activities, and hydrophobicity and for the presence of fimbriae. The presence of the 42-kDa (the fimbrilin subunit) and the 43- and 49-kDa (the HA-Ag2 components) polypeptides was investigated by immunoblotting using polyclonal and monoclonal antibodies directed to fimbriae and to the hemagglutinating adhesin HA-Ag2. Cells from two of the three mutants (M1 and M2) exhibited no or little hemagglutination activity and very low trypsin-like activity and did not show the 43- and 49-kDa polypeptides. Abnormal fimbriation in M1 was deduced from the following observations of cells grown for 18 h: absence of the 42-kDa polypeptide and of a 14-kDa polypeptide and no fimbriae visible on electron micrographs. While the cells of mutant M2, irrespective of the age of the culture, were found to lack the 43- and 49-kDa polypeptides and hemagglutination activity, the supernatants of cultures grown for 72 h had high hemagglutination and trypsin-like activities and revealed the presence of the 42-, 43-, and 49-kDa polypeptides. This suggests that M2 may be missing some molecules which anchor the components to the cell surface. Mutant M3 showed levels of activities similar to those of the parental strain but lacked the 43-kDa polypeptide. Other pleiotropic effects observed for the mutants included loss of dark pigmentation and lower hydrophobicity. The data from this study fuel an emerging consensus whereby fimbriation, hemagglutination, and proteolytic activities, as well as other functions in P. gingivalis, are intricate.
Chandad
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F
Groupe de Recherche en Ecologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, Canada.
Mayrand
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Grenier
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Hinode
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D
Mouton
C
C
D018829:Adhesins, Bacterial / Q000032:analysis; D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis; D010861:Fimbriae, Bacterial / Q000502:physiology; D006384:Hemagglutination; D037102:Lectins / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D009154:Mutation; D010641:Phenotype; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D011817:Rabbits
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D037102:Lectins