Immunological characterization and localization of a Porphyromonas gingivalis BApNA-hydrolyzing protease possessing hemagglutinating activity

A monoclonal antibody (mAb-PC) was produced against a BA pNA-hydrolyzing protease possessing hemagglutinating activity (Pase-C) from Porphyromonas gingivalis. Other P. gingivalis BA pNA-hydrolyzing enzymes (Pase-B and Pase-S) did not react with this antibody. By ELISA or SDS-PAGE and Western immunob...

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Bibliographic Details
Published in:FEMS microbiology letters 1995-09, Vol.131 (2), p.211-217
Main Authors: Hinode, D, Masuda, K, Yoshioka, M, Watanabe, K, Umemoto, T, Grenier, D, Mayrand, D, Nakamura, R
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal
Benzoylarginine Nitroanilide - metabolism
Cats
Dogs
Endopeptidases
Hemagglutination Inhibition Tests
Humans
Mice
Microscopy, Immunoelectron
Peptide Hydrolases - immunology
Peptide Hydrolases - metabolism
Porphyromonas - enzymology
Porphyromonas - isolation & purification
Porphyromonas gingivalis - enzymology
Porphyromonas gingivalis - isolation & purification
Porphyromonas gingivalis - ultrastructure
Species Specificity
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Summary:A monoclonal antibody (mAb-PC) was produced against a BA pNA-hydrolyzing protease possessing hemagglutinating activity (Pase-C) from Porphyromonas gingivalis. Other P. gingivalis BA pNA-hydrolyzing enzymes (Pase-B and Pase-S) did not react with this antibody. By ELISA or SDS-PAGE and Western immunoblotting analysis, mAb-PC recognized all P. gingivalis and P. endodontalis strains tested but did not recognize other members of the Porphyromonas genus nor other putative periodontopathogenic organisms. Pase-C, extracellular vesicles (ECV) and human strains of P. gingivalis showed two major immunoreactive bands (44 kDa and 40 kDa), whereas a different pattern was obtained with animal strains of P. gingivalis. Biotinylarginyl chloromethane, an irreversible inhibitor of trypsin-like proteases, did not affect the reactivity of Pase-C with mAb-PC on immunoblot. By reversed-phase electronmicroscopy following immunogold labeling, the antibody was shown to bind to the cell surface of P. gingivalis. mAb-PC inhibited the hemagglutinating activity of both P. gingivalis cells and ECV whereas a monoclonal antibody against LPS of P. gingivalis did not. These results suggest that Pase-C is located on the cell surface of P. gingivalis and may participate in erythrocyte binding.
ISSN:0378-1097
DOI:10.1016/0378-1097(95)00261-3