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Competitive Live-Cell Profiling Strategy for Discovering Inhibitors of the Quinolone Biosynthesis of Pseudomonas aeruginosa

Author:
Prothiwa, Michaela, Englmaier, Felix, Böttcher, Thomas
Source:
Journal of the American Chemical Society 2018 v.140 no.43 pp. 14019-14023
ISSN:
1520-5126
Subject:
Escherichia coli, Pseudomonas aeruginosa, animal pathogens, anthranilic acid, biosynthesis, enzymes, inhibitory concentration 50, models, quinolones, quorum sensing, virulence
Abstract:
Quinolones of the human pathogen Pseudomonas aeruginosa serve as antibacterial weapons and quorum sensing signals and coordinate the production of important virulence factors. A central enzyme for the biosynthesis of these quinolones is the synthetase PqsD. We developed an activity-based probe strategy that allows to screen for PqsD inhibitors in a cellular model system of live cells of Escherichia coli overexpressing PqsD. This strategy allowed us to determine IC₅₀ values for PqsD inhibition directly in live cells. Our most potent inhibitors were derived from the anthranilic acid core of the native substrate and resulted in single-digit micromolar IC₅₀ values. The effectiveness of our approach was ultimately demonstrated in P. aeruginosa by the complete shutdown of the production of quinolone quorum sensing signals and quinolone N-oxides and a considerable inhibition of the production of phenazine virulence factors.
Agid:
6194850