Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia. (1st January 2021)

• Record Type: Journal Article
• Authors: Zak, Krzysztof M.; Bostock, Mark J.; Waligorska, Irena; Thøgersen, Ida B.; Enghild, Jan J.; Popowicz, Grzegorz M.; Grudnik, Przemyslaw; Potempa, Jan; Ksiazek, Miroslaw
• Is Part Of: Journal of enzyme inhibition and medicinal chemistry. Volume 36:Number 1(2021)
• Page Start: 1267
• Page End: 1281
• Publication Date: 2021-01-01
• Subjects: Periodontitis -- proteolysis -- Tannerella forsythia -- NMR-based fragment screening -- protease inhibitors; Enzyme inhibitors -- Periodicals; Enzyme Inhibitors -- periodicals; Biochemistry -- periodicals; 572.7
• DOI: 10.1080/14756366.2021.1937619
• Languages: English
• ISSNs: 1475-6366

Abstract

Abstract: 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 ( K i = 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.