Functional fluorescent probe based on amide condensation of coumarin and melatonin for the turn-on detection of calcium ion in vitro and living cell

Abstract

Due to a physiological significant role of calcium ion, it is essential to develop ways of detecting and monitor the level of Ca2+ ions in living cell. Herein, we rationally designed and synthesized a functional blue fluorescent compound CM based on amide condensation of coumarin and melatonin. Interestingly, it could exhibit an “On-OFF” fluorescence response for Ca2+ in an aqueous environment with 80-fold enhancement in bright blue emission, which is also free from interference of other co-existing cations of high concentration. A detection limit of 0.28 ppm was obtained, and the binding ratio of CM and Ca2+ is further investigated by Job’s plot. After coordination of CM with calcium ion, a significant “Tyndall Effect” can be observed, and blue emissive nanowires induced by aggregation of complex of CM and Ca2+ were further investigated by Dynamic Light Scattering, analysis of Zeta potential, and confocal imaging. The molecular docking of CM with melatonin receptor 1A is also conducted. MTT assay reveals that CM ligand and the corresponding Ca2+ complex are non-cytotoxic. Moreover, CM shows good membrane permeability, and the relative cellular uptake mechanism is mainly dominated by an energy-dependent process. CM and its complex of Ca2+ can be further used as intracellular luminescent imaging probes and their localization in cell organelles is also revealed.