ArticlePloS one2011
Evaluation of multidrug efflux pump inhibitors by a new method using microfluidic channels.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 48 papers.
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Who cites it
48 citing papers in PubMed, 97 citations in OpenAlex.
- Review
- Quantifying bacterial efflux within subcellular domains ofApplied and environmental microbiology · 2024Article
- Mixed species biofilms act as planktonic cell factories despite isothiazolinone exposure under continuous-flow conditions.Environmental microbiology reports · 2024Article
- Investigation of the role of Cremophor RH 40 and Cremophor EL in the inhibition of efflux pump ofHeliyon · 2024Article
- Structural analysis of resistance-nodulation cell division transporters.Microbiology and molecular biology reviews : MMBR · 2024Review
- Scalable and Chromatography-Free Synthesis of Efflux Pump Inhibitor Phenylalanine Arginine β-Naphthylamide for Its Validation in Wild-Type Bacterial Strains.ChemMedChem · 2023Article
- Boosting the Antibacterial Activity of Azithromycin on Multidrug-ResistantInternational journal of molecular sciences · 2023Article
- Resuscitation dynamics reveal persister partitioning after antibiotic treatment.Molecular systems biology · 2023Article
- Membrane-Binding Biomolecules Influence the Rate of Vesicle Exchange between Bacteria.Applied and environmental microbiology · 2022Article
- An efflux-susceptible antibiotic-adjuvant with systemic efficacy against mouse infections.Scientific reports · 2022Article
- Structural and Functional Diversity of Resistance-Nodulation-Cell Division Transporters.Chemical reviews · 2021Review
- The Whole Is Bigger than the Sum of Its Parts: Drug Transport in the Context of Two Membranes with Active Efflux.Chemical reviews · 2021Article
- Plant-derived secondary metabolites as the main source of efflux pump inhibitors and methods for identification.Journal of pharmaceutical analysis · 2020Review
- Discovery of multidrug efflux pump inhibitors with a novel chemical scaffold.Biochimica et biophysica acta. General subjects · 2020Article
- Proof of an Outer Membrane Target of the Efflux Inhibitor Phe-Arg-β-Naphthylamide from Random Mutagenesis.Molecules (Basel, Switzerland) · 2019Article
- Polymyxin Derivatives that Sensitize Gram-Negative Bacteria to Other Antibiotics.Molecules (Basel, Switzerland) · 2019Review
- Characterization of β-lactamase- and efflux pump-mediated multiple antibiotic resistance inFood science and biotechnology · 2018Article
- A cell-based infection assay identifies efflux pump modulators that reduce bacterial intracellular load.PLoS pathogens · 2018Article
- Multiparametric Profiling for Identification of Chemosensitizers against Gram-Negative Bacteria.Frontiers in microbiology · 2018Article
- Loss of Methyltransferase Function and Increased Efflux Activity Leads to Doxycycline Resistance in Burkholderia pseudomallei.Antimicrobial agents and chemotherapy · 2017Article
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescein-di-β-D-galactopyranoside (FDG), a fluorogenic compound, is hydrolyzed by β-galactosidase in the cytoplasm of Escherichia coli to produce a fluorescent dye, fluorescein. We found that both FDG and fluorescein were substrates of efflux pumps, and have developed a new method to evaluate efflux-inhibitory activities in E. coli using FDG and a microfluidic channel device. We used E. coli MG1655 wild-type, ΔacrB (ΔB), ΔtolC (ΔC) and ΔacrBΔtolC (ΔBC) harboring plasmids carrying the mexAB-oprM (pABM) or mexXY-oprM (pXYM) genes of Pseudomonas aeruginosa. Two inhibitors, MexB-specific pyridopyrimidine (D13-9001) and non-specific Phe-Arg-β-naphthylamide (PAβN) were evaluated. The effects of inhibitors on pumps were observed using the microfluidic channel device under a fluorescence microscope. AcrAB-TolC and analogous pumps effectively prevented FDG influx in wild-type cells, resulting in no fluorescence. In contrast, ΔB or ΔC easily imported and hydrolyzed FDG to fluorescein, which was exported by residual pumps in ΔB. Consequently, fluorescent medium in ΔB and fluorescent cells of ΔC and ΔBC were observed in the microfluidic channels. D13-9001 substantially increased fluorescent cell number in ΔBC/pABM but not in ΔBC/pXYM. PAβN increased medium fluorescence in all strains, especially in the pump deletion mutants, and caused fluorescein accumulation to disappear in ΔC. The checkerboard method revealed that D13-9001 acts synergistically with aztreonam, ciprofloxacin, and erythromycin only against the MexAB-OprM producer (ΔBC/pABM), and PAβN acts synergistically, especially with erythromycin, in all strains including the pump deletion mutants. The results obtained from PAβN were similar to the results from membrane permeabilizer, polymyxin B or polymyxin B nonapeptide by concentration. The new method clarified that D13-9001 specifically inhibited MexAB-OprM in contrast to PAβN, which appeared to be a substrate of the pumps and permeabilized the membranes in E. coli.
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