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Bacterial Multidrug Exporters pp 71–81Cite as

Purification of AcrAB-TolC Multidrug Efflux Pump for Cryo-EM Analysis

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1700))

Abstract

The cell envelope of Gram-negative bacteria comprises an outer membrane, a cytoplasmic inner membrane, and an interstitial space. The tripartite multidrug transporter AcrAB-TolC, which uses proton electrochemical gradients to vectorially drive the efflux of drugs from the cell, spans this envelope. We describe here details of the methods used to prepare the recombinant tripartite assembly for high-resolution structure determination by cryo-EM.

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References

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Acknowledgments

Wellcome Trust and Human Frontier Science Program provided funding for this work. The work was also supported by NIH grant NIH 5P41GM103832.

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Authors and Affiliations

  1. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1GA, UK

    Dijun Du & Ben F. Luisi

  2. National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA

    Zhao Wang & Wah Chiu

Authors
  1. Dijun Du
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  2. Zhao Wang
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  3. Wah Chiu
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  4. Ben F. Luisi
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Corresponding author

Correspondence to Ben F. Luisi .

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Editors and Affiliations

  1. Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka, Japan

    Akihito Yamaguchi

  2. Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan

    Kunihiko Nishino

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Du, D., Wang, Z., Chiu, W., Luisi, B.F. (2018). Purification of AcrAB-TolC Multidrug Efflux Pump for Cryo-EM Analysis. In: Yamaguchi, A., Nishino, K. (eds) Bacterial Multidrug Exporters. Methods in Molecular Biology, vol 1700. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7454-2_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7454-2_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7452-8

  • Online ISBN: 978-1-4939-7454-2

  • eBook Packages: Springer Protocols

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