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Home / Publicações / Measuring Efflux and Permeability in Mycobacteria – chapter of «Mycobacteria Protocols»

Measuring Efflux and Permeability in Mycobacteria – chapter of «Mycobacteria Protocols»

  • Autores: Liliana Rodrigues, José A Aínsa, Miguel Viveiros
  • Ano de Publicação: 2021
  • Journal: Mycobacteria Protocols, 231-245, part of the book series, Methods in Molecular Biology, 2314
  • Link: https://doi.org/10.1007/978-1-0716-1460-0_9

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.

KEYWORDS

Accumulation assay; Efflux assay; Efflux inhibitors; Efflux pumps; Ethidium bromide; Fluorometry; Mycobacterium; Permeability.

Reference

Rodrigues L., Aínsa J.A., Viveiros M. (2021) Measuring Efflux and Permeability in Mycobacteria. In: Parish T., Kumar A. (eds) Mycobacteria Protocols, pp. 231-245. Methods in Molecular Biology, vol 2314. Humana, New York, NY.

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GHTM is a R&D Center that brings together researchers from IHMT with a track record in Tropical Medicine and International/Global Health. It aims at strengthening Portugal's role as a leading partner in the development and implementation of a global health research agenda. Our evidence-based interventions contribute to the promotion of equity in health and to improve the health of populations.

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