Novel mercapto fluorescence probe and uses thereof

Abstract

The invention belongs to the technical field of biological detection, which particularly relates to a fluorescent probe which can be applied in measuring active mercapto image in living cells and the application. The fluorescent probe is 2-(2-hydroxyphenyl) hydroxyphenyl compound which is decorated by maleimide or maleimide acid, the structure of the compound is N-{3-(2- benzimidazolyl)-4-hydroxyphenyl}maleimide and N-{3-(2- benzimidazolyl)-4-hydroxyphenyl}malemide acid. The compound can simply and rapidly get into living cells, which has a reaction with free active mercapto group in cells, and fluorescence intensity of the compound is led to be obviously enhancing, thereby the fluorescent probe can be used in fluorescent measurement or fluorescent image of fluactive mercapto group (such as glutathione and the like) in living cells.

Description

a technical field [0001] The invention belongs to the technical field of biological detection, specifically relates to a technique for measuring sulfhydryl content in living cells or living organisms, and is a fluorescent probe that can be used for imaging and measuring sulfhydryl in living cells and its application. Two background technology [0002] Sulfhydryl (-SH) is the most chemically active group in cells. In proteins, the sulfhydryl moiety is the most reactive functional group associated with enzymatic activity. The sulfhydryl group in cells, especially glutathione (GSH), as a nucleophilic and reducing agent in cells, can protect cells from hypoxia, toxins, mutagenesis, radiation and many carcinogens. With the gradual recognition of the important role of thiol functional groups in biological systems, methods for the identification and quantitative determination of biothiol compounds have attracted increasing attention. Traditional detection methods include nitropru...

AI Technical Summary

Problems solved by technology

[0004] The reported sulfhydryl fluorescent probes include aryl halides, dansylaziridines, etc., which have good selectivity and sensitivity, and the detection limit of sulfhydryl is generally around nanomolar (nmol), but these probes themselves All have strong fluorescence, which will cause a low signal-to-noise ratio when used in live cell imaging; there is also a class such as benzofuransulfonyl halide, although its autofluorescence is weak, but it needs to be used in alkaline conditions and above room temperature Under derivatization with sulfhydryl groups, it cannot be applied to live cell imaging

The most commonly used reagents for derivatizing sulfhydryl groups are acetyl halide derivatives, mainly iodoacetamide derivatives, which react quickly with sulfhydryl groups and can be carried out at room temperature and physiological pH, but the products are prone to lose fluorophores and are unstable to light

These existing sulfhydryl fluorescent probes cannot well meet the requirements of bioluminescence detection and fluorescence imaging.

[0005] Maleimide fluorescent compounds can undergo stable addition to sulfhydryl groups, and their autofluorescence is weak. After addition to sulfhydryl groups, the fluorescence increases. This characteristic of fluorescence enhancement only after the reaction makes it unique. Superiority, but after the general fluorophore is combined with maleimide, the quantum yield is low, the fluorescence lifetime is short, and the Stokes shift is small, it is difficult to perform high-quality intracellular fluorescence imaging

Images