Cytoplasmic Accumulation and Permeability of Antibiotics in Gram Positive and Gram Negative Bacteria Visualized in Real-Time via a Fluorogenic Tagging Strategy

被引：2
|
作者：
Squire, Scott O. [1 ,2 ]
Sebghati, Sepehr [1 ,2 ]
Hammond, Ming C. [1 ,2 ]
机构：
[1] Univ Utah, Dept Chem, Salt Lake City, UT 84112 USA
[2] Univ Utah, Henry Eyring Ctr Cell & Genome Sci, Salt Lake City, UT 84112 USA
来源：
ACS CHEMICAL BIOLOGY | 2023年 / 19卷 / 01期
基金：
美国国家卫生研究院; 美国国家科学基金会;
关键词：
OUTER-MEMBRANE; FLUORESCENT BIOSENSORS; DRUG ACCUMULATION; ASSAY; PENETRATION; RESISTANCE; BARRIER; PROBES;
D O I：
10.1021/acschembio.3c00510
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
In this study, we describe the first 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 bacteria.
引用
收藏
页码：3 / 8
页数：6
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