A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator.
To assess the direct effects of Bacteroides gingivalis on periodontal cells, human gingival fibroblasts were cultured in the presence of B. gingivalis extracts or a trypsinlike enzyme partially purified from the bacteria by chromatography on benzamidine-Sepharose and Sephacryl S-200. Analysis of cell surface glycoproteins by the periodate-[3H]borohydride labeling technique combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-fluorography demonstrated that fibronectin and some other high-molecular-weight cell surface glycoproteins were degraded by a 35,000-Mr(35K) B. gingivalis protease. Immunostaining of the fibroblast cultures showed degradation of intercellular matrix fibronectin by the 35K protease. The pattern of fibronectin degradation was monitored by examining the reaction products with the SDS-PAGE-immunoblotting technique. The protease degraded fibronectin rapidly and more extensively than did corresponding amounts of pancreatic trypsin. Collagenase secretion by the fibroblasts was assayed by incubating cell culture medium with soluble type I [3H]collagen at 25 degrees C followed by SDS-PAGE-fluorography analysis of the reaction products. The medium was also assayed for plasminogen activator activity by using a casein-agarose diffusion plate assay. The fibroblasts cultured with the 35K protease secreted increased amounts of collagenase and plasminogen activator into the medium. The results suggest that periodontal infection by B. gingivalis causes proteolytic damage of the host cell surface structures. Concomitantly, B. gingivalis may induce the cells to degrade their pericellular matrix.
Uitto
V J
VJ
Department of Oral Biology, University of British Columbia, Vancouver, Canada.
Larjava
H
H
Heino
J
J
Sorsa
T
T
D001439:Bacteroides / Q000201:enzymology* / Q000502:physiology; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000201:enzymology / Q000378:metabolism* / Q000382:microbiology; D005347:Fibroblasts / Q000166:cytology / Q000382:microbiology / Q000502:physiology; D005353:Fibronectins / Q000378:metabolism; D005881:Gingiva / Q000201:enzymology / Q000378:metabolism / Q000382:microbiology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism*; D003012:Microbial Collagenase / Q000378:metabolism*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000502:physiology*; D010960:Plasminogen Activators / Q000378:metabolism*
D005353:Fibronectins; D008562:Membrane Glycoproteins; D010447:Peptide Hydrolases; D010960:Plasminogen Activators; D003012:Microbial Collagenase