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Mycobacteria Protocols pp 231–245Cite as

Measuring Efflux and Permeability in Mycobacteria

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2314))

Abstract

Mycobacteria are intrinsically resistant to most antimicrobials, which is generally attributed to the impermeability of their cell wall that considerably limits drug uptake. Moreover, like in other pathogenic bacteria, active efflux systems have been widely characterized from diverse mycobacterial species in laboratory conditions, showing that they can promote resistance by extruding noxious compounds prior to their reaching their intended targets. Therefore, the intracellular concentration of a given compound is determined by the balance between permeability, influx, and efflux.

Given the urgent need to discover and develop novel antimycobacterial compounds in order to design effective therapeutic strategies, the contributions to drug resistance made by the controlled permeability of the cell wall and the increased activity of efflux pumps must be determined. In this chapter, we will describe a method that allows (1) the measuring of permeability and the quantification of general efflux activity of mycobacteria, by the study of the transport (influx and efflux) of fluorescent compounds, such as ethidium bromide; and (2) the screening of compounds in search of agents that increase the permeability of the cell wall and efflux inhibitors that could restore the effectiveness of antimicrobials that are subject to efflux.

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Acknowledgments

L. Rodrigues was supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 795924.

Author information

Authors and Affiliations

  1. Global Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, IHMT, Universidade Nova de Lisboa, UNL, Lisbon, Portugal

    Liliana Rodrigues & Miguel Viveiros

  2. Grupo de Genética de Micobacterias, Departamento de Microbiología, Pediatría, Radiología y Salud Pública, Facultad de Medicina, BIFI Universidad de Zaragoza-IIS Aragón, Zaragoza, Spain

    José A. Aínsa

  3. CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Madrid, Spain

    José A. Aínsa

Authors
  1. Liliana Rodrigues
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  2. José A. Aínsa
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  3. Miguel Viveiros
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Corresponding author

Correspondence to Liliana Rodrigues .

Editor information

Editors and Affiliations

  1. Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USA

    Tanya Parish

  2. Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USA

    Anuradha Kumar

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Rodrigues, L., Aínsa, J.A., Viveiros, M. (2021). Measuring Efflux and Permeability in Mycobacteria. In: Parish, T., Kumar, A. (eds) Mycobacteria Protocols. Methods in Molecular Biology, vol 2314. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1460-0_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1460-0_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1459-4

  • Online ISBN: 978-1-0716-1460-0

  • eBook Packages: Springer Protocols

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