Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia
Mirolysin is a secretory protease of Tannerella forsythia, a member of the dysbiotic oral microbiota responsible for periodontitis. In this study, we show that mirolysin latency is achieved by a “cysteine-switch” mechanism exerted by Cys23 in the N-terminal profragment. Mutation of Cys23 shortened t...
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Taylor & Francis Group
2021-01-01
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| Series: | Journal of Enzyme Inhibition and Medicinal Chemistry |
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| Online Access: | http://dx.doi.org/10.1080/14756366.2021.1937619 |
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doaj-facc41ee0c7f40019822708c69e05cfb2021-07-06T12:16:06ZengTaylor & Francis GroupJournal of Enzyme Inhibition and Medicinal Chemistry1475-63661475-63742021-01-013611267128110.1080/14756366.2021.19376191937619Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythiaKrzysztof M. Zak0Mark J. Bostock1Irena Waligorska2Ida B. Thøgersen3Jan J. Enghild4Grzegorz M. Popowicz5Przemyslaw Grudnik6Jan Potempa7Miroslaw Ksiazek8Helmholtz Zentrum München, Institute of Structural BiologyHelmholtz Zentrum München, Institute of Structural BiologyDepartment of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian UniversityDepartment of Molecular Biology and Genetics, Aarhus UniversityDepartment of Molecular Biology and Genetics, Aarhus UniversityHelmholtz Zentrum München, Institute of Structural BiologyMalopolska Centre of Biotechnology, Jagiellonian UniversityDepartment of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian UniversityDepartment of Microbiology, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian UniversityMirolysin is a secretory protease of Tannerella forsythia, a member of the dysbiotic oral microbiota responsible for periodontitis. In this study, we show that mirolysin latency is achieved by a “cysteine-switch” mechanism exerted by Cys23 in the N-terminal profragment. Mutation of Cys23 shortened the time needed for activation of the zymogen from several days to 5 min. The mutation also decreased the thermal stability and autoproteolysis resistance of promirolysin. Mature mirolysin is a thermophilic enzyme and shows optimal activity at 65 °C. Through NMR-based fragment screening, we identified a small molecule (compound (cpd) 9) that blocks promirolysin maturation and functions as a competitive inhibitor (Ki = 3.2 µM), binding to the S1′ subsite of the substrate-binding pocket. Cpd 9 shows superior specificity and does not interact with other T. forsythia proteases or Lys/Arg-specific proteases.http://dx.doi.org/10.1080/14756366.2021.1937619periodontitisproteolysistannerella forsythianmr-based fragment screeningprotease inhibitors |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Krzysztof M. Zak Mark J. Bostock Irena Waligorska Ida B. Thøgersen Jan J. Enghild Grzegorz M. Popowicz Przemyslaw Grudnik Jan Potempa Miroslaw Ksiazek |
| spellingShingle |
Krzysztof M. Zak Mark J. Bostock Irena Waligorska Ida B. Thøgersen Jan J. Enghild Grzegorz M. Popowicz Przemyslaw Grudnik Jan Potempa Miroslaw Ksiazek Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia Journal of Enzyme Inhibition and Medicinal Chemistry periodontitis proteolysis tannerella forsythia nmr-based fragment screening protease inhibitors |
| author_facet |
Krzysztof M. Zak Mark J. Bostock Irena Waligorska Ida B. Thøgersen Jan J. Enghild Grzegorz M. Popowicz Przemyslaw Grudnik Jan Potempa Miroslaw Ksiazek |
| author_sort |
Krzysztof M. Zak |
| title |
Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia |
| title_short |
Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia |
| title_full |
Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia |
| title_fullStr |
Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia |
| title_full_unstemmed |
Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia |
| title_sort |
latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen tannerella forsythia |
| publisher |
Taylor & Francis Group |
| series |
Journal of Enzyme Inhibition and Medicinal Chemistry |
| issn |
1475-6366 1475-6374 |
| publishDate |
2021-01-01 |
| description |
Mirolysin is a secretory protease of Tannerella forsythia, a member of the dysbiotic oral microbiota responsible for periodontitis. In this study, we show that mirolysin latency is achieved by a “cysteine-switch” mechanism exerted by Cys23 in the N-terminal profragment. Mutation of Cys23 shortened the time needed for activation of the zymogen from several days to 5 min. The mutation also decreased the thermal stability and autoproteolysis resistance of promirolysin. Mature mirolysin is a thermophilic enzyme and shows optimal activity at 65 °C. Through NMR-based fragment screening, we identified a small molecule (compound (cpd) 9) that blocks promirolysin maturation and functions as a competitive inhibitor (Ki = 3.2 µM), binding to the S1′ subsite of the substrate-binding pocket. Cpd 9 shows superior specificity and does not interact with other T. forsythia proteases or Lys/Arg-specific proteases. |
| topic |
periodontitis proteolysis tannerella forsythia nmr-based fragment screening protease inhibitors |
| url |
http://dx.doi.org/10.1080/14756366.2021.1937619 |
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