Discovery of Compounds Inhibiting the ADP-Ribosyltransferase Activity of Pertussis Toxin
| Autor: | Arto T. Pulliainen, Lari Lehtiö, Michael O. Hottiger, Yashwanth Ashok, Avlokita Tiwari, Moona Miettinen, Mark S. Johnson, Mahlet Z. Tamirat, Danilo Kimio Hirabae de Oliveira, Katja Näreoja |
|---|---|
| Přispěvatelé: | University of Zurich, Lehtiö, Lari |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Bordetella pertussis 030106 microbiology Molecular Dynamics Simulation Pertussis toxin Virulence factor Microbiology 03 medical and health sciences Inhibitory Concentration 50 Heterotrimeric G protein Drug Discovery medicine Escherichia coli Humans Secretion Whooping cough 030304 developmental biology G protein-coupled receptor ADP Ribose Transferases 0303 health sciences biology 030306 microbiology Chemistry 2725 Infectious Diseases Ligand (biochemistry) biology.organism_classification medicine.disease NAD 10226 Department of Molecular Mechanisms of Disease In vitro 3. Good health Molecular Docking Simulation 030104 developmental biology Infectious Diseases HEK293 Cells Biochemistry Pertussis Toxin ADP-ribosylation 570 Life sciences NAD+ kinase |
| Zdroj: | ACS infectious diseases. 6(4) |
| ISSN: | 2373-8227 |
| Popis: | Targeted pathogen-selective approach to antibiotic development holds promise to minimize collateral damage to the beneficial microbiome. The AB5-topology pertussis toxin (PtxS1-S5, 1:1:1:2:1) is a major virulence factor ofBordetella 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 mg-levels of truncated but highly active recombinantB. pertussisPtxS1 fromEscherichia coliand anin vitrohigh 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 thein vitroNAD+consumption assay that also were potent in an independentin vitroassay 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 AB5holotoxin-catalyzed ADP-ribosylation of Gαi in living human cells with a low micromolar IC50-value (2.4 µM). NSC228155 and NSC29193 might prove useful hit compounds in targetedB. pertussis-selective drug development. |
| Databáze: | OpenAIRE |
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