Measuring Efflux and Permeability in Mycobacteria

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages231-245
Number of pages15
Volume2314
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2314
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Accumulation assay
  • Efflux assay
  • Efflux inhibitors
  • Efflux pumps
  • Ethidium bromide
  • Fluorometry
  • Mycobacterium
  • Permeability

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