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In vitro transport activity of the fully assembled MexAB-OprM efflux pump from Pseudomonas aeruginosa

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  • Alice Verchère

    (Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie de Paris)

  • Manuela Dezi

    (Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie de Paris
    Present address: Institut de minéralogie, de physique des matériaux et de cosmochimie, UMR 7590, Université Pierre et Marie Curie, 4 place Jussieu, 75005 Paris, France.)

  • Vladimir Adrien

    (Laboratoire de physique statistique de l'École Normale Supérieure, UMR 8550, CNRS, Université Pierre et Marie Curie)

  • Isabelle Broutin

    (Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie de Paris)

  • Martin Picard

    (Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie de Paris)

Abstract

Antibiotic resistance is a major public health issue and many bacteria responsible for human infections have now developed a variety of antibiotic resistance mechanisms. For instance, Pseudomonas aeruginosa, a disease-causing Gram-negative bacteria, is now resistant to almost every class of antibiotics. Much of this resistance is attributable to multidrug efflux pumps, which are tripartite membrane protein complexes that span both membranes and actively expel antibiotics. Here we report an in vitro procedure to monitor transport by the tripartite MexAB-OprM pump. By combining proteoliposomes containing the MexAB and OprM portions of the complex, we are able to assay energy-dependent substrate translocation in a system that mimics the dual-membrane architecture of Gram-negative bacteria. This assay facilitates the study of pump transport dynamics and could be used to screen pump inhibitors with potential clinical use in restoring therapeutic activity of old antibiotics.

Suggested Citation

  • Alice Verchère & Manuela Dezi & Vladimir Adrien & Isabelle Broutin & Martin Picard, 2015. "In vitro transport activity of the fully assembled MexAB-OprM efflux pump from Pseudomonas aeruginosa," Nature Communications, Nature, vol. 6(1), pages 1-6, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms7890
    DOI: 10.1038/ncomms7890
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