Measuring Efflux and Permeability in Mycobacteria

Rodrigues, Liliana ; Aínsa, José A. (Universidad de Zaragoza) ; Viveiros, Miguel

Resumen: 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.

Idioma: Inglés

DOI: 10.1007/978-1-0716-1460-0_9

Año: 2021

Publicado en: Methods in Molecular Biology 2314 (2021), 231-245
ISSN: 1064-3745

Factor impacto CITESCORE: 2.5 - Biochemistry, Genetics and Molecular Biology (Q3)

Financiación: info:eu-repo/grantAgreement/EC/H2020/795924/EU/Chemogenomics and in silico repurposing as an innovative approach for rapid drug discovery in tuberculosis/iCHEMGENODRUGS_TB

Tipo y forma: Article (PostPrint)

Área (Departamento): Área Microbiología (Dpto. Microb.Ped.Radio.Sal.Pú.)