Tripartite assembly of RND multidrug efflux pumps

Laetitia Daury; François Orange; Jean-Christophe Taveau; Alice Verchère; Laura Monlezun; Céline Gounou; Ravi K R Marreddy; Martin Picard; Isabelle Broutin; Klaas M Pos; Olivier Lambert

doi:10.1038/ncomms10731

Tripartite assembly of RND multidrug efflux pumps

Nat Commun. 2016 Feb 12:7:10731. doi: 10.1038/ncomms10731.

Authors

Laetitia Daury^{1

2}, François Orange^{1

2}, Jean-Christophe Taveau^{1

2}, Alice Verchère³, Laura Monlezun³, Céline Gounou^{1

2}, Ravi K R Marreddy⁴, Martin Picard³, Isabelle Broutin³, Klaas M Pos⁴, Olivier Lambert^{1

2}

Affiliations

¹ Université de Bordeaux, CBMN UMR 5248, Bordeaux INP, IECB, Pessac F-33600, France.
² CNRS, CBMN UMR 5248, Pessac F-33600, France.
³ Laboratoire de Cristallographie et RMN Biologiques, UMR 8015, CNRS, Université Paris Descartes, Faculté de Pharmacie, 4 Avenue de l'Observatoire, Paris 75006, France.
⁴ Institute of Biochemistry, Goethe-University Frankfurt, Max-von-Laue-Str. 9, D-60438 Frankfurt am Main, Germany.

Abstract

Tripartite multidrug efflux systems of Gram-negative bacteria are composed of an inner membrane transporter, an outer membrane channel and a periplasmic adaptor protein. They are assumed to form ducts inside the periplasm facilitating drug exit across the outer membrane. Here we present the reconstitution of native Pseudomonas aeruginosa MexAB-OprM and Escherichia coli AcrAB-TolC tripartite Resistance Nodulation and cell Division (RND) efflux systems in a lipid nanodisc system. Single-particle analysis by electron microscopy reveals the inner and outer membrane protein components linked together via the periplasmic adaptor protein. This intrinsic ability of the native components to self-assemble also leads to the formation of a stable interspecies AcrA-MexB-TolC complex suggesting a common mechanism of tripartite assembly. Projection structures of all three complexes emphasize the role of the periplasmic adaptor protein as part of the exit duct with no physical interaction between the inner and outer membrane components.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Outer Membrane Proteins / metabolism*
Bacterial Outer Membrane Proteins / ultrastructure
Drug Resistance, Multiple, Bacterial*
Escherichia coli
Escherichia coli Proteins / metabolism*
Escherichia coli Proteins / ultrastructure
Lipoproteins / metabolism*
Lipoproteins / ultrastructure
Membrane Transport Proteins / metabolism*
Membrane Transport Proteins / ultrastructure
Microscopy, Electron, Transmission
Multidrug Resistance-Associated Proteins / metabolism*
Multidrug Resistance-Associated Proteins / ultrastructure
Multiprotein Complexes / metabolism*
Multiprotein Complexes / ultrastructure
Nanostructures
Native Polyacrylamide Gel Electrophoresis
Periplasmic Proteins / metabolism
Pseudomonas aeruginosa

Substances

AcrA protein, E coli
AcrB protein, E coli
Bacterial Outer Membrane Proteins
Escherichia coli Proteins
Lipoproteins
Membrane Transport Proteins
MexA protein, Pseudomonas aeruginosa
MexB protein, Pseudomonas aeruginosa
Multidrug Resistance-Associated Proteins
Multiprotein Complexes
OprM protein, Pseudomonas aeruginosa
Periplasmic Proteins
tolC protein, E coli