title: Gingipains of Porphyromonas gingivalis Affect the Stability and Function of Serine Protease Inhibitor of Kazal-type 6 (SPINK6), a Tissue Inhibitor of Human Kallikreins.
abstract: Periodontitis, a chronic inflammation driven by dysbiotic subgingival bacterial flora, is linked on clinical levels to the development of a number of systemic diseases and to the development of oral and gastric tract tumors. A key pathogen, Porphyromonas gingivalis, secretes gingipains, cysteine proteases implicated as the main factors in the development of periodontitis. Here we hypothesize that gingipains may be linked to systemic pathologies through the deregulation of kallikrein-like proteinase (KLK) family members. KLKs are implicated in cancer development and are clinically utilized as tumor progression markers. In tissues, KLK activity is strictly controlled by a limited number of tissue-specific inhibitors, including SPINK6, an inhibitor of these proteases in skin and oral epithelium. Here we identify gingipains as the only P. gingivalis proteases responsible for SPINK6 degradation. We further show that gingipains, even at low nanomolar concentrations, cleaved SPINK6 in concentration- and time-dependent manner. The proteolysis was accompanied by loss of inhibition against KLK13. We also mapped the cleavage by Arg-specific gingipains to the reactive site loop of the SPINK6 inhibitor. Moreover, we identified a significant fraction of SPINK6-sensitive proteases in healthy saliva and confirmed the ability of gingipains to inactivate SPINK6 under ex vivo conditions. Finally, we demonstrate the double-edge action of gingipains, which, in addition, can activate KLKs because of gingipain K-mediated proteolytic processing of the zymogenic proform of KLK13. Altogether, the results indicate the potential of P. gingivalis to disrupt the control system of KLKs, providing a possible mechanistic link between periodontal disease and tumor development.
authors.affiliation: From the Departments of Microbiology and.
authors.forename: Karolina
authors.initials: K
authors.lastname: Plaza
authors.affiliation: From the Departments of Microbiology and.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Kalinska
authors.affiliation: Analytical Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, and.
authors.forename: Oliwia
authors.initials: O
authors.lastname: Bochenska
authors.affiliation: the Department of Dermatology, University Clinic Schleswig-Holstein, 24105 Kiel, Germany, and.
authors.forename: Ulf
authors.initials: U
authors.lastname: Meyer-Hoffert
authors.affiliation: the Department of Dermatology, University Clinic Schleswig-Holstein, 24105 Kiel, Germany, and.
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Wu
authors.affiliation: the Department of Dermatology, University Clinic Schleswig-Holstein, 24105 Kiel, Germany, and.
authors.forename: Jan
authors.initials: J
authors.lastname: Fischer
authors.affiliation: From the Departments of Microbiology and.
authors.forename: Katherine
authors.initials: K
authors.lastname: Falkowski
authors.affiliation: From the Departments of Microbiology and.
authors.forename: Laura
authors.initials: L
authors.lastname: Sasiadek
authors.affiliation: From the Departments of Microbiology and.
authors.forename: Ewa
authors.initials: E
authors.lastname: Bielecka
authors.affiliation: the Department of Oral Immunology and Infectious Diseases, School of Dentistry, University of Louisville, Louisville, Kentucky 40202.
authors.forename: Barbara
authors.initials: B
authors.lastname: Potempa
authors.affiliation: Analytical Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, and.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Kozik
authors.affiliation: From the Departments of Microbiology and the Department of Oral Immunology and Infectious Diseases, School of Dentistry, University of Louisville, Louisville, Kentucky 40202 jan.potempa@uj.edu.pl.
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.affiliation: Malopolska Centre of Biotechnology, Jagiellonian University, 30-387 Krakow, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Kantyka
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D007610:Kallikreins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D055550:Protein Stability; D053491:Proteinase Inhibitory Proteins, Secretory / Q000737:chemistry / Q000378:metabolism*; D012463:Saliva / Q000737:chemistry / Q000378:metabolism*; D012471:Salivary Proteins and Peptides / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000074162:Serine Peptidase Inhibitors, Kazal Type
chemical_list: D018829:Adhesins, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D053491:Proteinase Inhibitory Proteins, Secretory; C553928:SPINK6 protein, human; D012471:Salivary Proteins and Peptides; D000074162:Serine Peptidase Inhibitors, Kazal Type; C470778:KLK13 protein, human; D007610:Kallikreins; D003546:Cysteine Endopeptidases
keywords: SPINK6; bacterial pathogenesis; enzyme inhibitor; gingipain; kallikrein; periodontal disease; protein degradation