Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement

Periodontitis is an inflammatory disease of the supporting structures of the teeth caused by, among other pathogens, Prevotella intermedia. Many strains of P. intermedia are resistant to killing by the human complement system, which is present at up to 70% of serum concentration in gingival crevicular fluid. Incubation of human serum with recombinant cysteine protease of P. intermedia (interpain A) resulted in a drastic decrease in bactericidal activity of the serum. Furthermore, a clinical strain 59 expressing interpain A was more serum-resistant than another clinical strain 57, which did not express interpain A, as determined by Western blotting. Moreover, in the presence of the cysteine protease inhibitor E64, the killing of strain 59 by human serum was enhanced. Importantly, we found that the majority of P. intermedia strains isolated from chronic and aggressive periodontitis carry and express the interpain A gene. The protective effect of interpain A against serum bactericidal activity was found to be attributable to its ability to inhibit all three complement pathways through the efficient degradation of the a-chain of C3—the major complement factor common to all three pathways. P. intermedia has been known to co-aggregate with P. gingivalis, which produce gingipains to efficiently degrade complement factors. Here, interpain A was found to have a synergistic effect with gingipains on complement degradation. In addition, interpain A was able to activate the C1 complex in serum, causing deposition of C1q on inert and bacterial surfaces, which may be important at initial stages of infection when local inflammatory reaction may be beneficial for a pathogen. Taken together, the newly characterized interpain A proteinase appears to be an important virulence factor of P. intermedia. Citation: Potempa M, Potempa J, Kantyka T, Nguyen K-A, Wawrzonek K, et al. (2009) Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement by Degrading Complement Factor C3. PLoS Pathog 5(2): e1000316. doi:10.1371/journal.ppat.1000316 Editor: Ulrich von Pawel-Rammingen, Umeå University, Sweden Received August 8, 2008; Accepted January 28, 2009; Published February 27, 2009 Copyright: 2009 Potempa et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by the Swedish Foundation for Strategic Research, the Swedish Research Council, Österlund Foundation, Kock Foundation, King Gustav Vth’s 80th Anniversary Foundation, Knut and Alice Wallenberg Foundation, Inga-Britt and Arne Lundberg Foundation, and research grants from the University Hospital in Malmö (to AB), and grants from the Ministry of Science and Higher Education (1642/B/P01/2008/35, Warsaw, Poland) and the National Institutes of Health (DE 09761, United States, to JP). Funding agencies had no influence in the design and conduct of the study, in the collection, analysis, and interpretation of the data, and in the preparation, review, or approval of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: [email protected]