Xu, Y, Song, S, Moeller, A, Kim, N, Piao, S, Sim, SH, Kang, M, Yu, W, Cho, HS, Chang, I, Lee, K and Ha, NC (2011) Functional implications of an intermeshing cogwheel-like interaction between TolC and MacA in the action of macrolide-specific efflux pump MacAB-TolC. J. Biol. Chem. 286:13541-9
Macrolide-specific efflux pump MacAB-TolC has been identified in diverse gram-negative bacteria including Escherichia coli. The inner membrane transporter MacB requires the outer membrane factor TolC and the periplasmic adaptor protein MacA to form a functional tripartite complex. In this study, we used a chimeric protein containing the tip region of the TolC α-barrel to investigate the role of the TolC α-barrel tip region with regard to its interaction with MacA. The chimeric protein formed a stable complex with MacA, and the complex formation was abolished by substitution at the functionally essential residues located at the MacA α-helical tip region. Electron microscopic study delineated that this complex was made by tip-to-tip interaction between the tip regions of the α-barrels of TolC and MacA, which correlated well with the TolC and MacA complex calculated by molecular dynamics. Taken together, our results demonstrate that the MacA hexamer interacts with TolC in a tip-to-tip manner, and implies the manner by which MacA induces opening of the TolC channel.
ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/metabolism; Actinobacillus actinomycetemcomitans/genetics; Actinobacillus actinomycetemcomitans/metabolism; Bacterial Outer Membrane Proteins/genetics; Bacterial Outer Membrane Proteins/metabolism; Biological Transport; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Macrolides/metabolism; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Multiprotein Complexes/genetics; Multiprotein Complexes/metabolism; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism
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