A palette of fluorophores that are differentially accumulated by wild- type and mutant strains of Escherichia coli: surrogate ligands for profiling bacterial membrane transporters

Salcedo-Sora, Jesus Enrique, Jindal, Srijan, O'Hagan, Steve and Kell, Douglas B ORCID: 0000-0001-5838-7963 (2021) A palette of fluorophores that are differentially accumulated by wild- type and mutant strains of Escherichia coli: surrogate ligands for profiling bacterial membrane transporters. MICROBIOLOGY-SGM, 167 (2). 001016-.

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Abstract

Our previous work demonstrated that two commonly used fluorescent dyes that were accumulated by wild-type <i>Escherichia coli</i> MG1655 were differentially transported in single-gene knockout strains, and also that they might be used as surrogates in flow cytometric transporter assays. We summarize the desirable properties of such stains, and here survey 143 candidate dyes. We eventually triage them (on the basis of signal, accumulation levels and cost) to a palette of 39 commercially available and affordable fluorophores that are accumulated significantly by wild-type cells of the 'Keio' strain BW25113, as measured flow cytometrically. Cheminformatic analyses indicate both their similarities and their (much more considerable) structural differences. We describe the effects of pH and of the efflux pump inhibitor chlorpromazine on the accumulation of the dyes. Even the 'wild-type' MG1655 and BW25113 strains can differ significantly in their ability to take up such dyes. We illustrate the highly differential uptake of our dyes into strains with particular lesions in, or overexpressed levels of, three particular transporters or transporter components (<i>yhjV</i>, <i>yihN</i> and <i>tolC</i>). The relatively small collection of dyes described offers a rapid, inexpensive, convenient and informative approach to the assessment of microbial physiology and phenotyping of membrane transporter function.

Item Type:	Article
Uncontrolled Keywords:	Douglas transporters, bioenergetics, E, coli, fluorescence, assays, energization
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User:	Symplectic Admin
Date Deposited:	08 Jul 2021 08:43
Last Modified:	15 Mar 2024 15:01
DOI:	10.1099/mic.0.001016
Open Access URL:	https://doi.org/10.1099/mic.0.001016
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3129254