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Discovery of Compounds Inhibiting the ADP-Ribosyltransferase Activity of Pertussis Toxin


Ashok, Yashwanth; Miettinen, Moona; De Oliveira, Danilo Kimio Hirabae; Tamirat, Mahlet Z; Näreoja, Katja; Tiwari, Avlokita; Hottiger, Michael O; Johnson, Mark S; Lehtiö, Lari; Pulliainen, Arto T (2020). Discovery of Compounds Inhibiting the ADP-Ribosyltransferase Activity of Pertussis Toxin. ACS Infectious Diseases, 6(4):588-602.

Abstract

The targeted pathogen-selective approach to drug development holds promise to minimize collateral damage to the beneficial microbiome. The AB$_{5}$-topology pertussis toxin (PtxS1-S5) is a major virulence factor of Bordetella pertussis, the causative agent of the highly contagious respiratory disease whooping cough. Once internalized into the host cell, PtxS1 ADP-ribosylates α-subunits of the heterotrimeric Gαi-superfamily, thereby disrupting G-protein-coupled receptor signaling. Here, we report the discovery of the first small molecules inhibiting the ADP-ribosyltransferase activity of pertussis toxin. We developed protocols to purify milligram-levels of active recombinant B. pertussis PtxS1 from Escherichia coli and an in vitro high throughput-compatible assay to quantify NAD$^{+}$ consumption during PtxS1-catalyzed ADP-ribosylation of Gαi. Two inhibitory compounds (NSC228155 and NSC29193) with low micromolar IC$_{50}$-values (3.0 μM and 6.8 μM) were identified in the in vitro NAD$^{+}$ consumption assay that also were potent in an independent in vitro assay monitoring conjugation of ADP-ribose to Gαi. Docking and molecular dynamics simulations identified plausible binding poses of NSC228155 and in particular of NSC29193, most likely owing to the rigidity of the latter ligand, at the NAD$^{+}$-binding pocket of PtxS1. NSC228155 inhibited the pertussis AB$_{5}$ holotoxin-catalyzed ADP-ribosylation of Gαi in living human cells with a low micromolar IC$_{50}$-value (2.4 μM). NSC228155 and NSC29193 might prove to be useful hit compounds in targeted B. pertussis-selective drug development.

Abstract

The targeted pathogen-selective approach to drug development holds promise to minimize collateral damage to the beneficial microbiome. The AB$_{5}$-topology pertussis toxin (PtxS1-S5) is a major virulence factor of Bordetella pertussis, the causative agent of the highly contagious respiratory disease whooping cough. Once internalized into the host cell, PtxS1 ADP-ribosylates α-subunits of the heterotrimeric Gαi-superfamily, thereby disrupting G-protein-coupled receptor signaling. Here, we report the discovery of the first small molecules inhibiting the ADP-ribosyltransferase activity of pertussis toxin. We developed protocols to purify milligram-levels of active recombinant B. pertussis PtxS1 from Escherichia coli and an in vitro high throughput-compatible assay to quantify NAD$^{+}$ consumption during PtxS1-catalyzed ADP-ribosylation of Gαi. Two inhibitory compounds (NSC228155 and NSC29193) with low micromolar IC$_{50}$-values (3.0 μM and 6.8 μM) were identified in the in vitro NAD$^{+}$ consumption assay that also were potent in an independent in vitro assay monitoring conjugation of ADP-ribose to Gαi. Docking and molecular dynamics simulations identified plausible binding poses of NSC228155 and in particular of NSC29193, most likely owing to the rigidity of the latter ligand, at the NAD$^{+}$-binding pocket of PtxS1. NSC228155 inhibited the pertussis AB$_{5}$ holotoxin-catalyzed ADP-ribosylation of Gαi in living human cells with a low micromolar IC$_{50}$-value (2.4 μM). NSC228155 and NSC29193 might prove to be useful hit compounds in targeted B. pertussis-selective drug development.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinärwissenschaftliches Institut > Department of Molecular Mechanisms of Disease
07 Faculty of Science > Department of Molecular Mechanisms of Disease
Dewey Decimal Classification:570 Life sciences; biology
Scopus Subject Areas:Health Sciences > Infectious Diseases
Language:English
Date:10 April 2020
Deposited On:15 Mar 2021 16:09
Last Modified:25 Jun 2024 01:38
Publisher:American Chemical Society (ACS)
ISSN:2373-8227
OA Status:Closed
Publisher DOI:https://doi.org/10.1021/acsinfecdis.9b00412
PubMed ID:31899865
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