title: A novel mechanism of latency in matrix metalloproteinases.
abstract: The matrix metalloproteinases (MMPs) are a family of secreted soluble or membrane-anchored multimodular peptidases regularly found in several paralogous copies in animals and plants, where they have multiple functions. The minimal consensus domain architecture comprises a signal peptide, a 60-90-residue globular prodomain with a conserved sequence motif including a cysteine engaged in "cysteine-switch" or "Velcro" mediated latency, and a catalytic domain. Karilysin, from the human periodontopathogen Tannerella forsythia, is the only bacterial MMP to have been characterized biochemically to date. It shares with eukaryotic forms the catalytic domain but none of the flanking domains. Instead of the consensus MMP prodomain, it features a 14-residue propeptide, the shortest reported for a metallopeptidase, which lacks cysteines. Here we determined the structure of a prokarilysin fragment encompassing the propeptide and the catalytic domain, and found that the former runs across the cleft in the opposite direction to a bound substrate and inhibits the latter through an "aspartate-switch" mechanism. This finding is reminiscent of latency maintenance in the otherwise unrelated astacin and fragilysin metallopeptidase families. In addition, in vivo and biochemical assays showed that the propeptide contributes to protein folding and stability. Our analysis of prokarilysin reveals a novel mechanism of latency and activation in MMPs. Finally, our findings support the view that the karilysin catalytic domain was co-opted by competent bacteria through horizontal gene transfer from a eukaryotic source, and later evolved in a specific bacterial environment.
authors.affiliation: From the Proteolysis Lab, Department of Structural Biology, Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, c/Baldiri Reixac, 15-21, 08028 Barcelona, Catalonia, Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: López-Pelegrín
authors.affiliation: the Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Ul. Gronostajowa 7, 30-387 Kraków, Poland, and.
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Ksiazek
authors.affiliation: the Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Ul. Gronostajowa 7, 30-387 Kraków, Poland, and.
authors.forename: Abdulkarim Y
authors.initials: AY
authors.lastname: Karim
authors.affiliation: From the Proteolysis Lab, Department of Structural Biology, Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, c/Baldiri Reixac, 15-21, 08028 Barcelona, Catalonia, Spain.
authors.forename: Tibisay
authors.initials: T
authors.lastname: Guevara
authors.affiliation: From the Proteolysis Lab, Department of Structural Biology, Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, c/Baldiri Reixac, 15-21, 08028 Barcelona, Catalonia, Spain,. Electronic address: jlacri@ibmb.csic.es.
authors.forename: Joan L
authors.initials: JL
authors.lastname: Arolas
authors.affiliation: the Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Ul. Gronostajowa 7, 30-387 Kraków, Poland, and; the Oral Immunology and Infectious Disease, University of Louisville School of Dentistry, Louisville, Kentucky 40202. Electronic address: jan.potempa@louisville.edu.
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
authors.affiliation: From the Proteolysis Lab, Department of Structural Biology, Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, c/Baldiri Reixac, 15-21, 08028 Barcelona, Catalonia, Spain,. Electronic address: fxgr@ibmb.csic.es.
authors.forename: F Xavier
authors.initials: FX
authors.lastname: Gomis-Rüth
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001438:Bacteroidaceae / Q000201:enzymology* / Q000235:genetics; D003545:Cysteine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019143:Evolution, Molecular; D022761:Gene Transfer, Horizontal; D006801:Humans; D020782:Matrix Metalloproteinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010518:Periodontitis / Q000201:enzymology / Q000235:genetics / Q000382:microbiology; D017510:Protein Folding; D017434:Protein Structure, Tertiary
chemical_list: D001426:Bacterial Proteins; D020782:Matrix Metalloproteinases; C572453:karilysin protein, Tannerella forsythia; D003545:Cysteine
keywords: Matrix Metalloproteinase (MMP); Peptidase; Periodontal Disease; Protease; Protein Evolution; X-ray Crystallography