Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan.
Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo, Kyoto 606-8501, Japan.
Org Biomol Chem. 2021 Oct 27;19(41):8906-8911. doi: 10.1039/d1ob01112j.
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 . We targeted a siderophore enterobactin biosynthetic enzyme EntE in . When coupled with an efflux pump inhibitor carbonyl cyanide -chlorophenylhydrazone, small-molecule probes were able to efficiently enter the cells, leading to the fluorescence labeling and imaging of overproduced EntE in . 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.
在荧光成像实验中，一个主要的挑战是小分子探针穿透细菌的外膜通透性屏障。本文描述了一种用于革兰氏阴性菌中荧光蛋白标记和成像的小分子探针的新策略。我们以铁载体 enterobactin 生物合成酶 EntE 为靶标。当与外排泵抑制剂羰基氰化物-对氯苯腙偶联时，小分子探针能够有效地进入细胞，导致. 中过量表达的 EntE 的荧光标记和成像。本研究表明，小分子探针与适当的外排泵抑制剂结合，可能会显著增强它们与活细菌中靶蛋白的相互作用。
