A cyanine dye-based fluorescent probe as indicator of copper clock reaction for tracing Cu²⁺-catalyzed oxidation of cysteine

Using cyanine dye-coupling 2-pyridylmethyl chelating unit as fluorescent probe, we successfully developed an indicator of copper clock reaction to monitor the process of cysteine (Cys) oxidation. During the monitoring assay, the fluorescent changes reflected three sequential reaction and binding behaviors including oxidation of Cys, oxidation of Cu(I) and binding between Cu(II) and the oxidation product cystine (Cys-Cys). Because the different kinetic trajectories were obtained when used Cys, GSH and Hcy as the substrates, respectively, a new strategy for distinguishing Cys from GSH and Hcy was further developed using the cyanine dye-coupling Cu(II) complex.