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

Krzysztof M Zak; Mark J Bostock; Irena Waligorska; Ida B Thøgersen; Jan J Enghild; Grzegorz M Popowicz; Przemyslaw Grudnik; Jan Potempa; Miroslaw Ksiazek

doi:10.1080/14756366.2021.1937619

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

Krzysztof M Zak, Mark J Bostock, Irena Waligorska, Ida B Thøgersen, Jan J Enghild, Grzegorz M Popowicz, Przemyslaw Grudnik, Jan Potempa, Miroslaw Ksiazek^*

^*Corresponding author for this work

Department of Molecular Biology and Genetics - Protein Science

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

3 Citations (Scopus)

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 (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.

Original language	English
Journal	Journal of Enzyme Inhibition and Medicinal Chemistry
Volume	36
Issue	1
Pages (from-to)	1267-1281
Number of pages	15
ISSN	1475-6366
DOIs	https://doi.org/10.1080/14756366.2021.1937619
Publication status	Published - 2021

Keywords

NMR-based fragment screening
Periodontitis
Tannerella forsythia
protease inhibitors
proteolysis
GINGIPAINS
ACTIVATION
MECHANISM
NMR-SPECTROSCOPY
PORPHYROMONAS-GINGIVALIS
PREVALENCE
SUPPRESSION
RED COMPLEX
VIRULENCE FACTORS
PERIODONTAL HEALTH

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Zak, K. M., Bostock, M. J., Waligorska, I., Thøgersen, I. B., Enghild, J. J., Popowicz, G. M., Grudnik, P., Potempa, J., & Ksiazek, M. (2021). 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, 36(1), 1267-1281. https://doi.org/10.1080/14756366.2021.1937619

@article{edb79c7387d5415fac5516d006fe8005,

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

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 (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.",

keywords = "NMR-based fragment screening, Periodontitis, Tannerella forsythia, protease inhibitors, proteolysis, GINGIPAINS, ACTIVATION, MECHANISM, NMR-SPECTROSCOPY, PORPHYROMONAS-GINGIVALIS, PREVALENCE, SUPPRESSION, RED COMPLEX, VIRULENCE FACTORS, PERIODONTAL HEALTH",

author = "Zak, {Krzysztof M} and Bostock, {Mark J} and Irena Waligorska and Th{\o}gersen, {Ida B} and Enghild, {Jan J} and Popowicz, {Grzegorz M} and Przemyslaw Grudnik and Jan Potempa and Miroslaw Ksiazek",

year = "2021",

doi = "10.1080/14756366.2021.1937619",

language = "English",

volume = "36",

pages = "1267--1281",

journal = "Journal of Enzyme Inhibition and Medicinal Chemistry",

issn = "1475-6366",

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Zak, KM, Bostock, MJ, Waligorska, I, Thøgersen, IB , Enghild, JJ, Popowicz, GM, Grudnik, P, Potempa, J & Ksiazek, M 2021, '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, vol. 36, no. 1, pp. 1267-1281. https://doi.org/10.1080/14756366.2021.1937619

Latency, thermal stability, and identification of an inhibitory compound of mirolysin, a secretory protease of the human periodontopathogen Tannerella forsythia. / Zak, Krzysztof M; Bostock, Mark J; Waligorska, Irena et al.
In: Journal of Enzyme Inhibition and Medicinal Chemistry, Vol. 36, No. 1, 2021, p. 1267-1281.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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 -

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

Abstract

Keywords

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