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Mirolase, a novel subtilisin-like serine protease from the periodontopathogen Tannerella forsythia

  • Miroslaw Ksiazek EMAIL logo , Abdulkarim Y. Karim , Danuta Bryzek , Jan J. Enghild , Ida B. Thøgersen , Joanna Koziel and Jan Potempa
Published/Copyright: November 13, 2014
Biological Chemistry
From the journal Biological Chemistry Volume 396 Issue 3

Abstract

The genome of Tannerella forsythia, an etiological factor of chronic periodontitis, contains several genes encoding putative proteases. Here, we characterized a subtilisin-like serine protease of T. forsythia referred to as mirolase. Recombinant full-length latent promirolase [85 kDa, without its signal peptide (SP)] processed itself through sequential autoproteolytic cleavages into a mature enzyme of 40 kDa. Mirolase latency was driven by the N-terminal prodomain (NTP). In stark contrast to almost all known subtilases, the cleaved NTP remained non-covalently associated with mirolase, inhibiting its proteolytic, but not amidolytic, activity. Full activity was observed only after the NTP was gradually, and fully, degraded. Both activity and processing was absolutely dependent on calcium ions, which were also essential for enzyme stability. As a consequence, both serine protease inhibitors and calcium ions chelators inhibited mirolase activity. Activity assays using an array of chromogenic substrates revealed that mirolase specificity is driven not only by the substrate-binding subsite S1, but also by other subsites. Taken together, mirolase is a calcium-dependent serine protease of the S8 family with the unique mechanism of activation that may contribute to T. forsythia pathogenicity by degradation of fibrinogen, hemoglobin, and the antimicrobial peptide LL-37.

Keywords: pathogenicity; periodontitis; protease specificity; protein expression; serine protease; virulence factor
Corresponding author: Miroslaw Ksiazek, Faculty of Biochemistry, Department of Microbiology, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387 Krakow, Poland, e-mail:

Acknowledgments

This study was supported by grants from the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University (K/DSC/000361), National Science Center, Poland (UMO-2011/03/N/NZ1/00586 and 2012/04/A/NZ1/00051 to M.K. and J.P., respectively), US NIH (DE 09761 and DE 022597), the European Commission (FP7-PEOPLE-2011-ITN-290246 ‘RAPID’ and FP7-HEALTH-F3-2012-306029 ‘TRIGGER’), and Polish Ministry of Science and Higher Education (UMO-2795/7.PR/13/2017/2). M.K. has obtained funding for the preparation of a doctoral dissertation from the National Science Center (Poland) as part of the funding of a doctoral scholarship on the basis of the decision’s number DEC-2013/08/T/NZ1/00315. The Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian University is a beneficiary of structural funds from the European Union (POIG.02.01.00-12-064/08).

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Received: 2014-10-2
Accepted: 2014-11-10
Published Online: 2014-11-13
Published in Print: 2015-3-1

©2015 by De Gruyter

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  2. Reviews
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  18. Cathepsin B-deficient mice as source of monoclonal anti-cathepsin B antibodies
https://doi.org/10.1515/hsz-2014-0256
Keywords for this article
pathogenicity; periodontitis; protease specificity; protein expression; serine protease; virulence factor

Articles in the same Issue

  1. Frontmatter
  2. Reviews
  3. The emerging role of the peptidome in biomarker discovery and degradome profiling
  4. Inflammatory caspases: key regulators of inflammation and cell death
  5. Minireview
  6. The role of serpins in tumor cell migration
  7. Research Articles/Short Communications
  8. Protein Structure and Function
  9. A core domain of the BMP2 proregion is sufficient for the biogenesis of mature homodimeric growth factor
  10. Molecular Medicine
  11. Role of nicotinamide N-methyltransferase in non-small cell lung cancer: in vitro effect of shRNA-mediated gene silencing on tumourigenicity
  12. Cell Biology and Signaling
  13. miR-103 promotes 3T3-L1 cell adipogenesis through AKT/mTOR signal pathway with its target being MEF2D
  14. miR-126 modulates angiogenic growth parameters of peripheral blood endothelial progenitor cells
  15. Osteoblastic alkaline phosphatase mRNA is stabilized by binding to vimentin intermediary filaments
  16. Proteolysis
  17. Mirolase, a novel subtilisin-like serine protease from the periodontopathogen Tannerella forsythia
  18. Cathepsin B-deficient mice as source of monoclonal anti-cathepsin B antibodies
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