Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates
Antibiotic-resistant Gram-negative bacteria are an increasing threat to human health. Strategies to restore antibiotic efficacy include targeting multidrug efflux pumps by competitive efflux pump inhibitors. These could be derived from natural substrates of these efflux systems. In this work, we aim...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1512472/full |
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| author | Léna Mazza Léna Mazza Alexandre Bory Alexandre Bory Alexandre Luscher Alexandre Luscher Joachim Kloehn Jean-Luc Wolfender Jean-Luc Wolfender Christian van Delden Christian van Delden Thilo Köhler Thilo Köhler |
| author_facet | Léna Mazza Léna Mazza Alexandre Bory Alexandre Bory Alexandre Luscher Alexandre Luscher Joachim Kloehn Jean-Luc Wolfender Jean-Luc Wolfender Christian van Delden Christian van Delden Thilo Köhler Thilo Köhler |
| author_sort | Léna Mazza |
| collection | DOAJ |
| description | Antibiotic-resistant Gram-negative bacteria are an increasing threat to human health. Strategies to restore antibiotic efficacy include targeting multidrug efflux pumps by competitive efflux pump inhibitors. These could be derived from natural substrates of these efflux systems. In this work, we aimed to elucidate the natural substrates of the clinically relevant Mex efflux pumps of Pseudomonas aeruginosa by an untargeted metabolomic approach. We constructed a PA14 mutant, genetically deleted in the major multidrug efflux pumps MexAB-OprM, MexCD-OprJ, MexXY-OprM, and MexEF-OprN and expressed in this mutant each efflux pump individually from an inducible promoter. Comparative analysis of the exo-metabolomes identified 210 features that were more abundant in the supernatant of efflux pump overexpressors compared to the pump-deficient mutant. Most of the identified features were efflux pump specific, while only a few were shared among several Mex pumps. We identified by-products of secondary metabolites as well as signaling molecules. Supernatants of the pump-deficient mutant also showed decreased accumulation of fatty acids, including long chain homoserine lactone quorum sensing molecules. Our data suggests that Mex efflux pumps of P. aeruginosa appear to have dedicated roles in extruding signaling molecules, metabolic by-products, as well as oxidized fatty acids. These findings represent an interesting starting point for the development of competitive efflux pump inhibitors. |
| format | Article |
| id | doaj-art-e32ad82370ec4d59b6790dc62eea55d0 |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-e32ad82370ec4d59b6790dc62eea55d02025-01-09T06:10:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011510.3389/fmicb.2024.15124721512472Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substratesLéna Mazza0Léna Mazza1Alexandre Bory2Alexandre Bory3Alexandre Luscher4Alexandre Luscher5Joachim Kloehn6Jean-Luc Wolfender7Jean-Luc Wolfender8Christian van Delden9Christian van Delden10Thilo Köhler11Thilo Köhler12Service of Infectious Diseases, Geneva University Hospitals, Geneva, SwitzerlandDepartment of Microbiology and Molecular Medicine, University of Geneva, Geneva, SwitzerlandSchool of Pharmaceutical Sciences, University of Geneva, Geneva, SwitzerlandInstitute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, SwitzerlandService of Infectious Diseases, Geneva University Hospitals, Geneva, SwitzerlandDepartment of Microbiology and Molecular Medicine, University of Geneva, Geneva, SwitzerlandDepartment of Microbiology and Molecular Medicine, University of Geneva, Geneva, SwitzerlandSchool of Pharmaceutical Sciences, University of Geneva, Geneva, SwitzerlandInstitute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, SwitzerlandService of Infectious Diseases, Geneva University Hospitals, Geneva, SwitzerlandDepartment of Microbiology and Molecular Medicine, University of Geneva, Geneva, SwitzerlandService of Infectious Diseases, Geneva University Hospitals, Geneva, SwitzerlandDepartment of Microbiology and Molecular Medicine, University of Geneva, Geneva, SwitzerlandAntibiotic-resistant Gram-negative bacteria are an increasing threat to human health. Strategies to restore antibiotic efficacy include targeting multidrug efflux pumps by competitive efflux pump inhibitors. These could be derived from natural substrates of these efflux systems. In this work, we aimed to elucidate the natural substrates of the clinically relevant Mex efflux pumps of Pseudomonas aeruginosa by an untargeted metabolomic approach. We constructed a PA14 mutant, genetically deleted in the major multidrug efflux pumps MexAB-OprM, MexCD-OprJ, MexXY-OprM, and MexEF-OprN and expressed in this mutant each efflux pump individually from an inducible promoter. Comparative analysis of the exo-metabolomes identified 210 features that were more abundant in the supernatant of efflux pump overexpressors compared to the pump-deficient mutant. Most of the identified features were efflux pump specific, while only a few were shared among several Mex pumps. We identified by-products of secondary metabolites as well as signaling molecules. Supernatants of the pump-deficient mutant also showed decreased accumulation of fatty acids, including long chain homoserine lactone quorum sensing molecules. Our data suggests that Mex efflux pumps of P. aeruginosa appear to have dedicated roles in extruding signaling molecules, metabolic by-products, as well as oxidized fatty acids. These findings represent an interesting starting point for the development of competitive efflux pump inhibitors.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1512472/fullPseudomonas aeruginosaantibiotic resistancemultidrug effluxmetabolomicsnatural substrates |
| spellingShingle | Léna Mazza Léna Mazza Alexandre Bory Alexandre Bory Alexandre Luscher Alexandre Luscher Joachim Kloehn Jean-Luc Wolfender Jean-Luc Wolfender Christian van Delden Christian van Delden Thilo Köhler Thilo Köhler Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates Frontiers in Microbiology Pseudomonas aeruginosa antibiotic resistance multidrug efflux metabolomics natural substrates |
| title | Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates |
| title_full | Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates |
| title_fullStr | Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates |
| title_full_unstemmed | Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates |
| title_short | Multidrug efflux pumps of Pseudomonas aeruginosa show selectivity for their natural substrates |
| title_sort | multidrug efflux pumps of pseudomonas aeruginosa show selectivity for their natural substrates |
| topic | Pseudomonas aeruginosa antibiotic resistance multidrug efflux metabolomics natural substrates |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1512472/full |
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