University of Oulu

Ashok, Y., Miettinen, M., Oliveira, D., Tamirat, M., Näreoja, K., Tiwari, A., Hottiger, M., Johnson, M., Lehtiö, L., Pulliainen, A. (2020) Discovery of Compounds Inhibiting the ADP-Ribosyltransferase Activity of Pertussis Toxin. ACS Infectious Diseases, 6 (4), 588-602, https://doi.org/10.1021/acsinfecdis.9b00412

Discovery of compounds inhibiting the ADP-ribosyltransferase activity of pertussis toxin

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Author: Ashok, Yashwanth1; Miettinen, Moona2; de Oliveira, Danilo Kimio Hirabae3;
Organizations: 1Faculty of Biochemistry and Molecular Medicine, Biocenter Oulu, University of Oulu, Aapistie 7A, P.O. Box 5400, FI-90014, Oulu, Finland
2Institute of Biomedicine, Research Centerfor Cancer, Infections, and Immunity and Turku DoctoralProgramme of Molecular Medicine (TuDMM), University ofTurku, FI-20520 Turku, Finland
3Faculty of Biochemistry and Molecular Medicine, Biocenter Oulu, University of Oulu, FI-90014 Oulu, Finland
4Structural Bioinformatics Laboratory, Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, FI-20520 Turku, Finland
5Institute of Biomedicine, Research Center for Cancer, Infections, and Immunity, University of Turku, FI-20520 Turku, Finland
6Department of Molecular Mechanisms of Disease, University of Zurich, 8057 Zurich, Switzerland;
Format: article
Version: accepted version
Access: embargoed
Persistent link: http://urn.fi/urn:nbn:fi-fe2020051435585
Language: English
Published: American Chemical Society, 2020
Publish Date: 2021-01-03
Description:

Abstract

The targeted pathogen-selective approach to drug development holds promise to minimize collateral damage to the beneficial microbiome. The AB₅-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 IC₅₀-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 AB₅ holotoxin-catalyzed ADP-ribosylation of Gαi in living human cells with a low micromolar IC₅₀-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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Series: ACS infectious diseases
ISSN: 2373-8227
ISSN-E: 2373-8227
ISSN-L: 2373-8227
Volume: 6
Issue: 4
Pages: 588 - 602
DOI: 10.1021/acsinfecdis.9b00412
OADOI: https://oadoi.org/10.1021/acsinfecdis.9b00412
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Funding: The research in the laboratory of A.T.P. is financially supported by Academy of Finland (grant no. 295296), Sigrid Juselius Foundation, Turku Doctoral Programme of Molecular Medicine (TuDMM) (to M.M.) and University of Turku, Turku, Finland. This work was funded in the L.L. laboratory by Academy of Finland (grant no. 287063, 294085, and 319299). The use of the facilities of the Biocenter Oulu for DNA sequencing, Proteomics and Protein Analysis and Protein Crystallography, a member of Biocenter Finland and Instruct-FI, is gratefully acknowledged. The laboratory of M.S.J. is supported by Sigrid Juselius Foundation, Joe, Pentti and Tor Memorial Fund, and Doctoral Network of Informational and Structural Biology (to M.T., Åbo Akademi Graduate School); computational infrastructure and core faculty support from Biocenter Finland (bioinformatics, structural biology and drug discovery and chemical biology nodes), CSC IT Center for Science; Academy of Finland FIRI infrastructure funding (grant no. 320005); screening core faculty of Biocity Turku, and Drug Discovery and Diagnostics strategic funding to Åbo Akademi University.
Academy of Finland Grant Number: 287063
294085
319299
Detailed Information: 287063 (Academy of Finland Funding decision)
294085 (Academy of Finland Funding decision)
319299 (Academy of Finland Funding decision)
Copyright information: This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Infectious Diseases, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see [insert ACS Articles on Request author-directed link to Published Work, see https://doi.org/10.1021/acsinfecdis.9b00412.