Cytoplasmic accumulation and permeability of antibiotics in Gram positive and Gram negative bacteria visualized in real-time via a fluorogenic tagging strategy

20 March 2023, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

In this study, we describe a real-time live cell assay for compound accumulation and permeability in both Gram positive and Gram negative bacteria. The assay utilizes a novel fluorogenic tagging strategy that permits direct visualization of compound accumulation dynamics in the cytoplasm of live cells, unobscured by washing or other processing steps. Quantitative differences could be reproducibly measured by flow cytometry at compound concentrations below the limit of detection for MS-based approaches. We establish the fluorogenic assay in E. coli and B. subtilis and compare the intracellular accumulation of two antibiotics, ciprofloxacin and ampicillin, with related pharmacophores in these model Gram negative and Gram positive bacteria.

Keywords

Fluorogenic antibiotics
RNA aptamer
Fluorescence

Supplementary materials

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Supporting Information for Cytoplasmic accumulation and permeability of antibiotics in Gram positive and Gram negative bacteria visualized in real-time via a fluorogenic tagging strategy
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