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Biochemical Reconstitution and Characterization of Multicomponent Drug Efflux Transporters

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Bacterial Multidrug Exporters

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1700))

Abstract

Efflux pumps are the major determinants in bacterial multidrug resistance. In Gram-negative bacteria, efflux transporters are organized as macromolecular tripartite machineries that span the two-membrane cell envelope of the bacterium. Biochemical data on purified proteins are essential to draw a mechanistic picture of this highly dynamical, multicomponent, efflux system. We describe protocols for the reconstitution and the in vitro study of transporters belonging to RND and ABC superfamilies: the AcrAB–TolC and MacAB–TolC efflux systems from Escherichia coli and the MexAB–OprM efflux pump from Pseudomonas aeruginosa.

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Author information

Authors and Affiliations

  1. Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie de Paris, 4 Avenue de l’Observatoire, 75006, Paris, France

    Isabelle Broutin & Alice Verchère

  2. Department of Chemistry and Biochemistry, University of Oklahoma, 101 Stephenson Parkway, Norman, OK, 73019, USA

    Shuo Lu & Helen I. Zgurskaya

  3. Department of Cell Biology and Biochemistry, Texas Tech University Health Science Center, 3601 4th Street, Lubbock, TX, 79430, USA

    Elena B. Tikhonova

  4. Laboratoire de Biologie Physico-chimique des ProtÕines Membranaires, UMR7099. IBPC, UniversitÕ Paris Diderot, 13, Rue Pierre et Marie Curie, 75005, Paris, France

    Martin Picard

Authors
  1. Martin Picard
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  2. Elena B. Tikhonova
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  3. Isabelle Broutin
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  4. Shuo Lu
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  5. Alice Verchère
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  6. Helen I. Zgurskaya
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Corresponding authors

Correspondence to Martin Picard or Helen I. Zgurskaya .

Editor information

Editors and Affiliations

  1. Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka, Japan

    Akihito Yamaguchi

  2. Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan

    Kunihiko Nishino

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Picard, M., Tikhonova, E.B., Broutin, I., Lu, S., Verchère, A., Zgurskaya, H.I. (2018). Biochemical Reconstitution and Characterization of Multicomponent Drug Efflux Transporters. In: Yamaguchi, A., Nishino, K. (eds) Bacterial Multidrug Exporters. Methods in Molecular Biology, vol 1700. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7454-2_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7454-2_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7452-8

  • Online ISBN: 978-1-4939-7454-2

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