Lighting‐Up of the Dye Malachite Green with Mercury(II)–DNA and Its Application for Fluorescence Turn‐Off Detection of Cysteine and Glutathione

Abstract

This work describes a novel strategy for the highly sensitive and selective detection of cysteine (Cys) and glutathione (GSH) based on the Hg(2+)-AGRO100-malachite green (MG) complex system. The dye MG, which has a very low quantum yield in aqueous solution by itself, can bind with the thymine-rich DNA AGRO100 in the presence of Hg(2+) ions to generate a striking fluorescence intensity enhancement of 1000-fold. As sulfur-containing amino acids, Cys and GSH effectively sequester Hg(2+) ions from the Hg(2+)-AGRO100-MG complex structure to switch the 'lit-up' chemosensor to the 'off' state (about a 50-fold fluorescence intensity decrease), thus providing a facile, but effective, method to probe for Cys/GSH. The fluorescence titration, UV absorption, CD, and Raman spectra provide some insight into the structural and chemical basis for the enhancement effect. The formation of the Hg(2+)-AGRO100-MG complex significantly affects the electronic structure and conformation of the MG molecule by leading to an extended π system, which is the likely origin of the observed striking fluorescence intensity enhancement. Notably, the proposed sensing platform exhibits exquisite selectivity and sensitivity toward Cys/GSH with limits of detection of 5 nM for Cys and 10 nM for GSH, respectively. Furthermore, the straightforward assay design avoids labeling of the probe, uses only commercially available materials, and still displays comparable sensitivity and excellent selectivity.