title: Cleavage of extracellular matrix in periodontitis: gingipains differentially affect cell adhesion activities of fibronectin and tenascin-C.
abstract: Gingipains are cysteine proteases that represent major virulence factors of the periodontopathogenic bacterium Porphyromonas gingivalis. Gingipains are reported to degrade extracellular matrix (ECM) of periodontal tissues, leading to tissue destruction and apoptosis. The exact mechanism is not known, however. Fibronectin and tenascin-C are pericellular ECM glycoproteins present in periodontal tissues. Whereas fibronectin mediates fibroblast adhesion, tenascin-C binds to fibronectin and inhibits its cell-spreading activity. Using purified proteins in vitro, we asked whether fibronectin and tenascin-C are cleaved by gingipains at clinically relevant concentrations, and how fragmentation by the bacterial proteases affects their biological activity in cell adhesion. Fibronectin was cleaved into distinct fragments by all three gingipains; however, only arginine-specific HRgpA and RgpB but not lysine-specific Kgp destroyed its cell-spreading activity. This result was confirmed with recombinant cell-binding domain of fibronectin. Of the two major tenascin-C splice variants, the large but not the small was a substrate for gingipains, indicating that cleavage occurred primarily in the alternatively spliced domain. Surprisingly, cleavage of large tenascin-C variant by all three gingipains generated fragments with increased anti-adhesive activity towards intact fibronectin. Fibronectin and tenascin-C fragments were detected in gingival crevicular fluid of a subset of periodontitis patients. We conclude that cleavage by gingipains directly affects the biological activity of both fibronectin and tenascin-C in a manner that might lead to increased cell detachment and loss during periodontal disease.
authors: Department of Orthodontics and Dentofacial Orthopedics, University of Bern, Switzerland.
authors: Sabrina
authors: S
authors: Ruggiero
authors: Raluca
authors: R
authors: Cosgarea
authors: Jan
authors: J
authors: Potempa
authors: Barbara
authors: B
authors: Potempa
authors: Sigrun
authors: S
authors: Eick
authors: Matthias
authors: M
authors: Chiquet
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism; D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D001120:Arginine / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D008239:Lysine / Q000378:metabolism; D051379:Mice; D010518:Periodontitis / Q000235:genetics / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000472:pathogenicity; D011485:Protein Binding; D019063:Tenascin / Q000235:genetics / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D005353:Fibronectins; D000080867:Gingipain Cysteine Endopeptidases; D019063:Tenascin; D001120:Arginine; D003546:Cysteine Endopeptidases; D008239:Lysine