Inhibition of efflux pumps aids small-molecule probe-based fluorescence labeling and imaging in the Gram-negative bacterium Escherichia coli. Issue 41 (31st August 2021)

• Record Type: Journal Article
• Authors: Ishikawa, Fumihiro; Konno, Sho; Takashima, Katsuki; Kakeya, Hideaki; Tanabe, Genzoh
• Is Part Of: Organic & biomolecular chemistry. Volume 19:Issue 41(2021)
• Page Start: 8906
• Page End: 8911
• Publication Date: 2021-08-31
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/d1ob01112j
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : We describe a novel strategy for small-molecule probe-based fluorescence protein labeling and imaging in the Gram-negative bacterium Escherichia coli, which involves inhibition of efflux pumps. Abstract : A major challenge in fluorescence imaging experiments, which are essential to determine protein activity, expression, and localization, is the penetration of small-molecule probes through the outer membrane permeability barrier of bacteria. Here, we describe a novel strategy for small-molecule probe-based fluorescence protein labeling and imaging in the Gram-negative bacterium Escherichia coli . We targeted a siderophore enterobactin biosynthetic enzyme EntE in E. coli . When coupled with an efflux pump inhibitor carbonyl cyanide m -chlorophenylhydrazone, small-molecule probes were able to efficiently enter the cells, leading to the fluorescence labeling and imaging of overproduced EntE in E. coli . This study demonstrates that the combination of small-molecule probes with appropriate efflux pump inhibitors may substantially enhance their interaction with the target proteins in live bacteria.