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Updated: Feb 18, 2026

A Proteoliposome-Based Efflux Assay to Determine Single-molecule Properties of Cl- Channels and Transporters
Published on: April 20, 2015
Biochemical Reconstitution and Characterization of Multicomponent Drug Efflux Transporters
Martin Picard1, Elena B Tikhonova2, Isabelle Broutin3
1Laboratoire de Biologie Physico-chimique des ProtÕines Membranaires, UMR7099. IBPC, UniversitÕ Paris Diderot, 13, Rue Pierre et Marie Curie, 75005, Paris, France. martin.picard@ibpc.fr.
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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