Crystal Structure of the Membrane Fusion Protein, MexA, of the Multidrug Transporter in Pseudomonas aeruginosa

Akama, Hiroyuki; Matsuura, Takanori; Kashiwagi, Sachiko; Yoneyama, Hiroshi; Narita, Shin‐ichiro; Tsukihara, Tomitake; Nakagawa, Atsushi; Nakae, Taiji

doi:10.1074/jbc.c400164200

Cited by 231 publications

(217 citation statements)

References 39 publications

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“…S7 for the multidrug resistance protein MexA of Pseudomonas aeruginosa, where nine subunits oligomerize to form a funnel-like structure across the periplasmic space for antibiotic efflux (PDB ID 1VF7; ref. 34).…”

mentioning

confidence: 99%

Direct-coupling analysis of residue coevolution captures native contacts across many protein families

Morcos

Pagnani

Lunt

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

1,356

2,225

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The similarity in the three-dimensional structures of homologous proteins imposes strong constraints on their sequence variability. It has long been suggested that the resulting correlations among amino acid compositions at different sequence positions can be exploited to infer spatial contacts within the tertiary protein structure. Crucial to this inference is the ability to disentangle direct and indirect correlations, as accomplished by the recently introduced direct-coupling analysis (DCA). Here we develop a computationally efficient implementation of DCA, which allows us to evaluate the accuracy of contact prediction by DCA for a large number of protein domains, based purely on sequence information. DCA is shown to yield a large number of correctly predicted contacts, recapitulating the global structure of the contact map for the majority of the protein domains examined. Furthermore, our analysis captures clear signals beyond intradomain residue contacts, arising, e.g., from alternative protein conformations, ligand-mediated residue couplings, and interdomain interactions in protein oligomers. Our findings suggest that contacts predicted by DCA can be used as a reliable guide to facilitate computational predictions of alternative protein conformations, protein complex formation, and even the de novo prediction of protein domain structures, contingent on the existence of a large number of homologous sequences which are being rapidly made available due to advances in genome sequencing.statistical sequence analysis | residue-residue covariation | contact map prediction | maximum-entropy modeling

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mentioning

confidence: 99%

Direct-coupling analysis of residue coevolution captures native contacts across many protein families

Morcos

Pagnani

Lunt

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

1,356

2,225

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“…These interactions can be detected by cross-linking in vivo and measured in vitro by isothermal calorimetry (ITC) (6). Adaptor monomers have an elongated modular structure comprising a ␤-barrel, a lipoyl domain, and a long ␣-helical hairpin that extends over four or five heptad repeats and projects into the periplasm (18)(19)(20).…”

mentioning

confidence: 99%

A periplasmic coiled-coil interface underlying TolC recruitment and the assembly of bacterial drug efflux pumps

Lobedanz

Bokma²,

Symmons³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

120

156

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Bacteria such as Escherichia coli and Pseudomonas aeruginosa expel antibiotics and other inhibitors via tripartite multidrug efflux pumps spanning the inner and outer membranes and the intervening periplasmic space. A key event in pump assembly is the recruitment of an outer membrane-anchored TolC exit duct by the adaptor protein of a cognate inner membrane translocase, establishing a contiguous transenvelope efflux pore. We describe the underlying interaction of juxtaposed periplasmic exit duct and adaptor coiled-coils in the widespread RND-type pump TolC/AcrAB of E. coli, using in vivo cross-linking to map the extent of intermolecular contacts. Cross-linking of site-specific TolC cysteine variants to wild-type AcrA adaptor identified residues on the lower ␣-helical barrel domain of TolC, defining a contiguous cluster close to the entrance aperture of the exit duct. Reciprocally, site-specific cross-linking of AcrA cysteine variants to wild-type TolC identified the interaction surface on the adaptor within the N-terminal ␣-helix of the AcrA coiled-coil. The experimental data allowed a data-driven docking approach to model the interaction surface central to pump assembly. The lowest energy docked model satisfying all of the cross-link distance constraints places the adaptor at the intramolecular groove formed by the TolC entrance helices, aligning the adaptor coiled-coil with the exposed TolC outer helix. A key feature of this positioning is that it allows space for the proposed movement of the inner coil of TolC during transition to its open state.antibiotic resistance ͉ exit duct ͉ membrane proteins ͉ type I export

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“…In 2000, sequencing of the genome of P. aeruginosa led to the identification of at least 12 efflux pumps 2 . The constitutively expressed tripartite efflux pump consists of the following: MexB, a protein from the resistance-nodulation-division family (RND) located within the inner membrane that binds the antibiotic and initiates its transport using the proton motive force 3 ; MexA, a membrane fusion protein (MFP), attached to the inner membrane with a lipid anchor, found in the periplasmic space, that is essential for in vivo transport and pump assembly 4,5 ; and OprM, a channel from the outer membrane factor family (OMF) located in the outer membrane that allows the extrusion of antibiotics 6 . High-resolution structures have been described for the various components [3][4][5][6] and shed light on the molecular determinants of transport.…”

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confidence: 99%

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

et al. 2015

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Crystal Structure of the Membrane Fusion Protein, MexA, of the Multidrug Transporter in Pseudomonas aeruginosa

Cited by 231 publications

References 39 publications

Direct-coupling analysis of residue coevolution captures native contacts across many protein families

Direct-coupling analysis of residue coevolution captures native contacts across many protein families

A periplasmic coiled-coil interface underlying TolC recruitment and the assembly of bacterial drug efflux pumps

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

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