Efflux transporters of the RND family confer resistance to multiple antibiotics in Gram-negative bacteria. Here, we identify and chemically optimize pyridylpiperazine-based compounds that potentiate antibiotic activity in E. coli through inhibition of its primary RND transporter, AcrAB-TolC. Characterisation of resistant E. coli mutants and structural biology analyses indicate that the compounds bind to a unique site on the transmembrane domain of the AcrB L protomer, lined by key catalytic residues involved in proton relay. Molecular dynamics simulations suggest that the inhibitors access this binding pocket from the cytoplasm via a channel exclusively present in the AcrB L protomer. Thus, our work unveils a class of allosteric efflux-pump inhibitors that likely act by preventing the functional catalytic cycle of the RND pump.

Pyridylpiperazine-based allosteric inhibitors of RND-type multidrug efflux pumps

Malloci G.;Vargiu A. V.;
2022-01-01

Abstract

Efflux transporters of the RND family confer resistance to multiple antibiotics in Gram-negative bacteria. Here, we identify and chemically optimize pyridylpiperazine-based compounds that potentiate antibiotic activity in E. coli through inhibition of its primary RND transporter, AcrAB-TolC. Characterisation of resistant E. coli mutants and structural biology analyses indicate that the compounds bind to a unique site on the transmembrane domain of the AcrB L protomer, lined by key catalytic residues involved in proton relay. Molecular dynamics simulations suggest that the inhibitors access this binding pocket from the cytoplasm via a channel exclusively present in the AcrB L protomer. Thus, our work unveils a class of allosteric efflux-pump inhibitors that likely act by preventing the functional catalytic cycle of the RND pump.
2022
Allosteric Regulation; Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Biological Transport; Crystallography, X-Ray; Drug Resistance, Multiple, Bacterial; Escherichia coli; Escherichia coli Proteins; Gene Expression; Lipoproteins; Membrane Transport Proteins; Molecular Dynamics Simulation; Multidrug Resistance-Associated Proteins; Mutation; Oligopeptides; Oxacillin; Piperazines; Promoter Regions, Genetic; Protein Binding; Protein Conformation, alpha-Helical; Protein Conformation, beta-Strand; Protein Interaction Domains and Motifs; Pyridines; Structure-Activity Relationship
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11584/330689
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