000117945 001__ 117945 000117945 005__ 20220817102228.0 000117945 0247_ $$2doi$$a10.1007/978-1-0716-1460-0_9 000117945 0248_ $$2sideral$$a129259 000117945 037__ $$aART-2021-129259 000117945 041__ $$aeng 000117945 100__ $$aRodrigues, Liliana 000117945 245__ $$aMeasuring Efflux and Permeability in Mycobacteria 000117945 260__ $$c2021 000117945 5060_ $$aAccess copy available to the general public$$fUnrestricted 000117945 5203_ $$aMycobacteria 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. 000117945 536__ $$9info:eu-repo/grantAgreement/EC/H2020/795924/EU/Chemogenomics and in silico repurposing as an innovative approach for rapid drug discovery in tuberculosis/iCHEMGENODRUGS_TB$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 795924-iCHEMGENODRUGS_TB 000117945 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/ 000117945 594__ $$a2.5$$b2021 000117945 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion 000117945 700__ $$0(orcid)0000-0003-2076-844X$$aAínsa, José A.$$uUniversidad de Zaragoza 000117945 700__ $$aViveiros, Miguel 000117945 7102_ $$11011$$2630$$aUniversidad de Zaragoza$$bDpto. Microb.Ped.Radio.Sal.Pú.$$cÁrea Microbiología 000117945 773__ $$g2314 (2021), 231-245$$pMethods mol. biol. (Clifton N.J.)$$tMethods in Molecular Biology$$x1064-3745 000117945 8564_ $$s360213$$uhttps://zaguan.unizar.es/record/117945/files/texto_completo.pdf$$yPostprint 000117945 8564_ $$s671486$$uhttps://zaguan.unizar.es/record/117945/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint 000117945 909CO $$ooai:zaguan.unizar.es:117945$$particulos$$pdriver 000117945 951__ $$a2022-08-17-09:16:46 000117945 980__ $$aARTICLE