Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia

TY - JOUR

T1 - Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia

AU - Zak, Krzysztof M

AU - Bostock, Mark J

AU - Waligorska, Irena

AU - Thøgersen, Ida B

AU - Enghild, Jan J

AU - Popowicz, Grzegorz M

AU - Grudnik, Przemyslaw

AU - Potempa, Jan

AU - Ksiazek, Miroslaw

PY - 2021

Y1 - 2021

N2 - 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.

AB - 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.

KW - NMR-based fragment screening

KW - Periodontitis

KW - Tannerella forsythia

KW - protease inhibitors

KW - proteolysis

KW - GINGIPAINS

KW - ACTIVATION

KW - MECHANISM

KW - NMR-SPECTROSCOPY

KW - PORPHYROMONAS-GINGIVALIS

KW - PREVALENCE

KW - SUPPRESSION

KW - RED COMPLEX

KW - VIRULENCE FACTORS

KW - PERIODONTAL HEALTH

U2 - 10.1080/14756366.2021.1937619

DO - 10.1080/14756366.2021.1937619

M3 - Journal article

C2 - 34210221

SN - 1475-6366

VL - 36

SP - 1267

EP - 1281

JO - Journal of Enzyme Inhibition and Medicinal Chemistry

JF - Journal of Enzyme Inhibition and Medicinal Chemistry

IS - 1

ER -