Tripartite efflux pumps: energy is required for dissociation, but not assembly or opening of the outer membrane channel of the pump

Article


Janganan T.K., Bavro, V.N., Zhang, L., Borges-Walmsley, M.I. and Walmsley, A.R. 2013. Tripartite efflux pumps: energy is required for dissociation, but not assembly or opening of the outer membrane channel of the pump. Molecular Microbiology. 88 (3), pp. 590-602. https://doi.org/10.1111/mmi.12211
TypeArticle
TitleTripartite efflux pumps: energy is required for dissociation, but not assembly or opening of the outer membrane channel of the pump
AuthorsJanganan T.K., Bavro, V.N., Zhang, L., Borges-Walmsley, M.I. and Walmsley, A.R.
Abstract

The MtrCDE multidrug pump, from Neisseria gonorrhoeae, is assembled from the inner and outer membrane proteins MtrD and MtrE, which are connected by the periplasmic membrane fusion protein MtrC. Although it is clear that MtrD delivers drugs to the channel of MtrE, it remains unclear how drug delivery and channel opening are connected. We used a vancomycin sensitivity assay to test for opening of the MtrE channel. Cells expressing MtrE or MtrE-E434K were insensitive to vancomycin; but became moderately and highly sensitive to vancomycin respectively, when coexpressed with MtrC, suggesting that the MtrE channel opening requires MtrC binding and is energy-independent. Cells expressing wild-type MtrD, in an MtrCE background, were vancomycin-insensitive, but moderately sensitive in an MtrCE-E434K background. The mutation of residues involved in proton translocation inactivated MtrD and abolished drug efflux, rendered both MtrE and MtrE-E434K vancomycin-insensitive; imply that the pump–component interactions are preserved, and that the complex is stable in the absence of proton flux, thus sealing the open end of MtrE. Following the energy-dependent dissociation of the tripartite complex, the MtrE channel is able to reseal, while MtrE-E434K is unable to do so, resulting in the vancomycin-sensitive phenotype. Thus, our findings suggest that opening of the OMP via interaction with the MFP is energy-independent, while both drug export and complex dissociation require active proton flux.

Sustainable Development Goals3 Good health and well-being
Middlesex University ThemeHealth & Wellbeing
PublisherWiley
JournalMolecular Microbiology
ISSN0950-382X
Electronic1365-2958
Publication dates
Online09 Apr 2013
Print22 Apr 2013
Publication process dates
Accepted19 Mar 2013
Deposited12 Nov 2024
Output statusPublished
Publisher's version
License
File Access Level
Open
Copyright Statement

© 2013 The Authors. Molecular Microbiology Published by Blackwell Publishing Ltd

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Digital Object Identifier (DOI)https://doi.org/10.1111/mmi.12211
Permalink -

https://repository.mdx.ac.uk/item/1w65y1

  • 2
    total views
  • 0
    total downloads
  • 0
    views this month
  • 0
    downloads this month

Export as

Related outputs

Genome sequence of the biocontrol agent Coniothyrium minitans Conio (IMI 134523)
Patel, D., Shittu, T.A., Baroncelli, R., Muthumeenakshi, S., Osborne, T.H., Janganan, T. and Sreenivasaprasad, S. 2021. Genome sequence of the biocontrol agent Coniothyrium minitans Conio (IMI 134523). Molecular Plant-Microbe Interactions. 34 (2), pp. 222-225. https://doi.org/10.1094/MPMI-05-20-0124-A
Architecture and self-assembly of Clostridium sporogenes and Clostridium botulinum spore surfaces illustrate a general protective strategy across spore formers
Janganan, T.K.,, Mullin, N., Dafis-Sagarmendi, A., Brunt, J., Tzokov, S.B., Stringer, S., Moir, A., Chaudhuri, R.R., Fagan, R.P., Hobbs, J.K. and Bullough, P.A. 2020. Architecture and self-assembly of Clostridium sporogenes and Clostridium botulinum spore surfaces illustrate a general protective strategy across spore formers. mSphere. 5 (4). https://doi.org/10.1128/msphere.00424-20
ATP-binding cassette transporter VcaM from Vibrio cholerae is dependent on the outer membrane factor family for its function
Lu, W.J., Lin, H.J., Janganan, T.K., Li, C.Y., Chin, W.C., Bavro, V.N. and Lin, H.T.V. 2018. ATP-binding cassette transporter VcaM from Vibrio cholerae is dependent on the outer membrane factor family for its function. International Journal of Molecular Sciences. 19 (4). https://doi.org/10.3390/ijms19041000
Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains
Janganan, T.K., Mullin, N., Stringer, S., Fagan, R.P., Hobbs, J.K., Moir, A. and Bullough, P.A. 2016. Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains. Food Microbiology. 59, pp. 205-212. https://doi.org/10.1016/j.fm.2016.06.003
A Gβ protein and the TupA co-regulator bind to protein kinase a Tpk2 to act as antagonistic molecular switches of fungal morphological changes
Janganan T.K., Chen, G., Chen, D., Menino, J.F., Rodrigues, F., Borges-Walmsley, M.I. and Walmsley, A.R. 2015. A Gβ protein and the TupA co-regulator bind to protein kinase a Tpk2 to act as antagonistic molecular switches of fungal morphological changes. PLoS ONE. 10 (9). https://doi.org/10.1371/journal.pone.0136866
Opening of the outer membrane protein channel in tripartite efflux pumps is induced by interaction with the membrane fusion partner
Janganan, T.K., Zhang, L., Barrera, N.P., Bavro, V.N., Vinkovic, D.M., Robinson, C.V., Borges-Walmsley, M.I. and Walmsley, A.R. 2011. Opening of the outer membrane protein channel in tripartite efflux pumps is induced by interaction with the membrane fusion partner. Journal of Biological Chemistry. 286 (7), pp. 5484-5493. https://doi.org/10.1074/jbc.M110.187658
Evidence for the assembly of a bacterial tripartite multidrug pump with a stoichiometry of 3: 6: 3
Janganan T.K., Bavro, V.N., Zhang, L., Vinkovic, D.M., Barrera, N.P., Robinson, C.V., Borges-Walmsley, M.I. and Walmsley, A.R. 2011. Evidence for the assembly of a bacterial tripartite multidrug pump with a stoichiometry of 3: 6: 3. Journal of Biological Chemistry. 286 (30), pp. 26900-26912. https://doi.org/10.1074/jbc.M111.246595Als
The regulation of the CAMP signalling pathway in the human pathogenic fungus, Paracoccidioides brasiliensis
Janganan, T.K. 2008. The regulation of the CAMP signalling pathway in the human pathogenic fungus, Paracoccidioides brasiliensis. PhD thesis Durham University School of Biosciences
The cAMP pathway is important for controlling the morphological switch to the pathogenic yeast form of Paracoccidioides brasiliensis
Chen, D., Janganan, T.K., Chen, G., Marques, E.R., Kress, M.R., Goldman, G.H., Walmsley, A.R. and Borges-Walmsley, M.I. 2007. The cAMP pathway is important for controlling the morphological switch to the pathogenic yeast form of Paracoccidioides brasiliensis. Molecular Microbiology. 65 (3), pp. 761-779. https://doi.org/10.1111/j.1365-2958.2007.05824.x