Effect of the proton motive force inhibitor carbonyl cyanide-<i>m</i>-chlorophenylhydrazone (CCCP) on<i>Pseudomonas aeruginosa</i>biofilm development

Ikonomidis, Alexandros; Tsakris, Athanassios; Kanellopoulou, Maria; Maniatis, Antonios N.; Pournaras, Spyros

doi:10.1111/j.1472-765x.2008.02430.x

Cited by 52 publications

(25 citation statements)

References 19 publications

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“…In the presence of CCCP (pump inhibitor), a single green NP (17 nm) stayed inside the cells for 75.0 min, much longer than 18.3 min observed in the absence of CCCP. The CCCP decreases or eliminates proton concentration gradients across the cellular membranes of living cells, disabling the proton-motive transporters (e.g., MexAB-OprM) and leading to the slow efflux rate (31, 55). Therefore, the results suggest that the NPs are indeed extruded by the proton-motive pumps.…”

Section: Resultsmentioning

confidence: 99%

Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosa Cells Using Size-Dependent Plasmonic Nanoparticle Optical Probes

et al. 2010

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Multidrug membrane transporters (efflux pumps) in both prokaryotes and eukaryotes are responsible for impossible treatments of a wide variety of diseases, including infections and cancer, underscoring the importance of better understanding their structures and functions for design of effective therapies. In this study, we designed and synthesized two silver nanoparticle (Ag NP) solution with average diameters of 13.1 ± 2.5 nm (8.1–38.6 nm) and 91.0 ± 9.3 nm (56–120 nm), and used the size-dependent plasmonic spectra of single NPs to probe the size-dependent transport kinetics of MexAB-OprM (multidrug transporter) in Pseudomonas (P.) aeruginosa in real-time at nanometer resolution. We found that the accumulation of intracellular NPs in wild-type (WT) cells was higher than in nalB1 (over-expression of MexAB-OprM), but less than ΔABM (deletion of MexAB-OprM). In the presence of proton ionophores (CCCP, inhibitor of proton-motive-force), we found that intracellular NPs in nalB1 were nearly doubled. These results suggest that the MexAB-OprM is responsible for the extrusion of NPs out of cells and NPs (orders of magnitude larger than conventional antibiotics) are the substrates of the transporter, which indicates that the substrates may trigger the assembly of the efflux pump optimized for the extrusion of the encountered substrates. We found that the smaller NPs stayed longer inside the cells than larger NPs, suggesting the size-dependent efflux kinetics of the cells. This study shows that multi-sized NPs can be used to mimic various sizes of antibiotics for probing the size-dependent efflux kinetics of multidrug membrane transporters in single living cells.

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Section: Resultsmentioning

confidence: 99%

Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosa Cells Using Size-Dependent Plasmonic Nanoparticle Optical Probes

et al. 2010

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“…For the P11C plate (antibiotics), there was no appreciable growth of any strain after 10 days of incubation. The compounds that showed the higher growth differences like benzoate and carbonyl cyanide-mchlorophenylhydrazone (CCCP), are uncouplers of the oxidative phosphorylation, and at least for CCCP its uncoupling activity has been demonstrated for P. aeruginosa (Ikonomidis et al, 2008). Azide is an inhibitor of complex IV of the respiratory chain, the enzyme cytochrome oxidase, and menadione is a compound that produces oxidative stress (Smirnova et al, 2000;Criddle et al, 2006).…”

Section: Resultsmentioning

confidence: 99%

Quorum sensing enhancement of the stress response promotes resistance to quorum quenching and prevents social cheating

et al. 2014

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Quorum sensing (QS) coordinates the expression of virulence factors and allows bacteria to counteract the immune response, partly by increasing their tolerance to the oxidative stress generated by immune cells. Despite the recognized role of QS in enhancing the oxidative stress response, the consequences of this relationship for the bacterial ecology remain unexplored. Here we demonstrate that QS increases resistance also to osmotic, thermal and heavy metal stress. Furthermore a QS-deficient lasR rhlR mutant is unable to exert a robust response against H 2 O 2 as it has less induction of catalase and NADPH-producing dehydrogenases. Phenotypic microarrays revealed that the mutant is very sensitive to several toxic compounds. As the anti-oxidative enzymes are private goods not shared by the population, only the individuals that produce them benefit from their action. Based on this premise, we show that in mixed populations of wild-type and the mexR mutant (resistant to the QS inhibitor furanone C-30), treatment with C-30 and H 2 O 2 increases the proportion of mexR mutants; hence, oxidative stress selects resistance to QS compounds. In addition, oxidative stress alone strongly selects for strains with active QS systems that are able to exert a robust anti oxidative response and thereby decreases the proportion of QS cheaters in cultures that are otherwise prone to invasion by cheats. As in natural environments stress is omnipresent, it is likely that this QS enhancement of stress tolerance allows cells to counteract QS inhibition and invasions by social cheaters, therefore having a broad impact in bacterial ecology.

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“…In this study, multidrug resistance due to overexpression of the efflux systems was evaluated by phenotypic tests in the presence of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) as an efflux pump inhibitor. CCCP is a known proton motive force and efflux pump inhibitor [8] that can be added in MuellerHinton agar during its preparation. The MICs of antibiotics for strains overexpressing efflux pumps are usually twofold or higher than those strains of species that do not overexpress efflux pumps.…”

Section: Introductionmentioning

confidence: 99%

Role of efflux pump inhibitor in decreasing antibiotic cross-resistance of Pseudomonas aeruginosa in a burn hospital in Iran

Talebi-Taher

Majidpour

Gholami

et al. 2016

J Infect Dev Ctries

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Introduction: Multidrug resistance in Pseudomonas aeruginosa may be due to efflux pump overexpression. This study phenotypically examined the role of efflux pump inhibitors in decreasing antibiotic cross-resistance between beta-lactams, fluoroquinolones, and aminoglycosides in P. aeruginosa isolates from burn patients in Iran. Methodology: A total of 91 phenotypically and genotypically confirmed P. aeruginosa samples were studied. Multidrug cross-resistance was determined using the disk diffusion method and minimum inhibitory concentration (MIC) test. The contribution of efflux pumps was determined by investigating MIC reduction assay to markers of beta-lactams, fluoroquinolones, and aminoglycosides in the absence and presence of an efflux pump inhibitor. All the isolates were also tested by polymerase chain reaction for the presence of mexA, mexC, and mexE efflux genes. Results: Of the isolates, 81 (89%) and 83 (91.2%) were multidrug resistant according to the disk diffusion and MIC method, respectively. Cross-resistance was observed in 67 (73.6%) and 68 (74.7%) of isolates according to the disk diffusion and MIC method, respectively. In the presence of the efflux pump inhibitor, twofold or higher MIC reduction to imipenem, cefepime, ciprofloxacin, and gentamicin was observed in 59, 65, 55, and 60 isolates, respectively. Except for two isolates that were negative for mexC, all isolates were positive for mexA, mexC, and mexE genes simultaneously. Conclusion: Efflux pumps could cause different levels of resistance based on their expression in clinical isolates. Early detection of different efflux pumps in P. aeruginosa could allow the use of other antibiotics and efflux pump inhibitors in combination with antibiotic therapy.

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Effect of the proton motive force inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP) onPseudomonas aeruginosabiofilm development

Cited by 52 publications

References 19 publications

Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosa Cells Using Size-Dependent Plasmonic Nanoparticle Optical Probes

Study of the Multidrug Membrane Transporter of Single Living Pseudomonas aeruginosa Cells Using Size-Dependent Plasmonic Nanoparticle Optical Probes

Quorum sensing enhancement of the stress response promotes resistance to quorum quenching and prevents social cheating

Role of efflux pump inhibitor in decreasing antibiotic cross-resistance of Pseudomonas aeruginosa in a burn hospital in Iran

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