Ligand‐transporter interaction in the AcrB multidrug efflux pump determined by fluorescence polarization assay
Open Access
- 25 September 2007
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 581 (25), 4972-4976
- https://doi.org/10.1016/j.febslet.2007.09.035
Abstract
The AcrB of Escherichia coli pumps out a wide range of compounds, including most of the currently available antibiotics, and contributes significantly to the serious problem of multidrug resistance of pathogenic bacteria. Quantitative analysis of drug efflux by this pump requires the measurement of the affinity of ligands. Yet there has been no success in determining these values. We introduce here an approach of steady‐state fluorescence polarization to study the interactions between four different ligands and the purified AcrB transporter in a detergent environment. Our assays indicate that the transporter binds these drugs with values ranging from 5.5 to 74.1 μM.This publication has 30 references indexed in Scilit:
- Characterization of the multidrug efflux regulator AcrR from Escherichia coliBiochemical and Biophysical Research Communications, 2007
- Crystal structure of the multidrug efflux transporter AcrB at 3.1Å resolution reveals the N-terminal region with conserved amino acidsJournal of Structural Biology, 2007
- Multidrug-Binding Transcription Factor QacR Binds the Bivalent Aromatic Diamidines DB75 and DB359 in Multiple PositionsJournal of the American Chemical Society, 2007
- Drug Export Pathway of Multidrug Exporter AcrB Revealed by DARPin InhibitorsPLoS Biology, 2006
- Conformation of the AcrB Multidrug Efflux Pump in Mutants of the Putative Proton Relay PathwayJournal of Bacteriology, 2006
- Threonine-978 in the Transmembrane Segment of the Multidrug Efflux Pump AcrB of Escherichia coli Is Crucial for Drug Transport as a Probable Component of the Proton Relay NetworkJournal of Bacteriology, 2006
- Structural Asymmetry of AcrB Trimer Suggests a Peristaltic Pump MechanismScience, 2006
- A Periplasmic Drug-Binding Site of the AcrB Multidrug Efflux Pump: a Crystallographic and Site-Directed Mutagenesis StudyJournal of Bacteriology, 2005
- Structural Basis of Multiple Drug-Binding Capacity of the AcrB Multidrug Efflux PumpScience, 2003
- Application of fluorescence energy transfer and polarization to monitor Escherichia coli cAMP receptor protein and lac promoter interaction.Proceedings of the National Academy of Sciences of the United States of America, 1990