Structure-based mechanism of cysteine-switch latency and of catalysis by pappalysin-family metallopeptidases
| Autor: | Miroslaw Ksiazek, Arturo Rodríguez-Banqueri, Jan Potempa, F.X. Gomis-Ruth, Tibisay Guevara |
|---|---|
| Přispěvatelé: | Generalitat de Catalunya, National Institutes of Health (US), Polish Academy of Sciences, Ministerio de Economía y Competitividad (España), Fundació La Marató de TV3, Ministry of Science and Higher Education (Poland) |
| Rok vydání: | 2020 |
| Předmět: |
Peridontopathogens
Metallopeptidase mirolysin catalytic mechanisms Cleavage (embryo) pappalysin family Biochemistry 03 medical and health sciences zymogens Zymogens Zymogen Hydrolase Mirolysin Tannerella forsythia General Materials Science Methanosarcina acetivorans Metallopeptidases lcsh:Science 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology peridontopathogens Catalytic mechanisms 030302 biochemistry & molecular biology General Chemistry Condensed Matter Physics biology.organism_classification Research Papers metallopeptidases Pappalysin family Enzyme chemistry metallopeptidases lcsh:Q Cysteine |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname IUCrJ, Vol 7, Iss 1, Pp 18-29 (2020) Digital.CSIC: Repositorio Institucional del CSIC Consejo Superior de Investigaciones Científicas (CSIC) IUCrJ |
| ISSN: | 2015-6448 |
| Popis: | Tannerella forsythia is an oral dysbiotic periodontopathogen involved in severe human periodontal disease. As part of its virulence factor armamentarium, at the site of colonization it secretes mirolysin, a metallopeptidase of the unicellular pappalysin family, as a zymogen that is proteolytically auto-activated extracellularly at the Ser54–Arg55 bond. Crystal structures of the catalytically impaired promirolysin point mutant E225A at 1.4 and 1.6 Å revealed that latency is exerted by an N-terminal 34-residue pro-segment that shields the front surface of the 274-residue catalytic domain, thus preventing substrate access. The catalytic domain conforms to the metzincin clan of metallopeptidases and contains a double calcium site, which acts as a calcium switch for activity. The pro-segment traverses the active-site cleft in the opposite direction to the substrate, which precludes its cleavage. It is anchored to the mature enzyme through residue Arg21, which intrudes into the specificity pocket in cleft sub-site S1′. Moreover, residue Cys23 within a conserved cysteine–glycine motif blocks the catalytic zinc ion by a cysteine-switch mechanism, first described for mammalian matrix metallopeptidases. In addition, a 1.5 Å structure was obtained for a complex of mature mirolysin and a tetradecapeptide, which filled the cleft from sub-site S1′ to S6′. A citrate molecule in S1 completed a product-complex mimic that unveiled the mechanism of substrate binding and cleavage by mirolysin, the catalytic domain of which was already preformed in the zymogen. These results, including a preference for cleavage before basic residues, are likely to be valid for other unicellular pappalysins derived from archaea, bacteria, cyanobacteria, algae and fungi, including archetypal ulilysin from Methanosarcina acetivorans. They may further apply, at least in part, to the multi-domain orthologues of higher organisms. This study was supported in part by grants from Spanish, Catalan, US American (NIH/NIDR) and Polish (NCN) public agencies (BFU2015-64487R; MDM-2014-0435; Fundacio´ ‘La Marato´ de TV3’ 201815 and 2017SGR3, 2015/17/B/NZ1/ 00666, 2016/21/B/NZ1/00292, and R21DE026280). MK was recipient of a scholarship from the Polish Ministry of Science and Higher Education (1306/MOB/IV/2015/0, ‘Mobilnoc´ Plus’). The Structural Biology Unit of IBMB was a ‘Marı´a de Maeztu’ Unit of Excellence of the Spanish Ministry of Science, Innovation and Universities (2015–2019). |
| Databáze: | OpenAIRE |
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