A bifunctional peptide-based fluorescent probe for ratiometric and “turn-on” detection of Zn(II) ions and its application in living cells

In this work, a bifunctional peptide-based fluorescent probe L containing a tetrapeptide scaffold (Pro-Gly-His-Trp-NH2) and a dansyl group was synthesized using solid phase peptide synthesis (SPPS) technology. As designed, L, based on a FRET mechanism, exhibited high selectivity, excellent ratiometric signals, and fast response to Zn2+ in aqueous solutions at an excitation wavelength of 280 nm. In addition, when excited at 320 nm, L exhibited a fluorescent "turn-on" response towards Zn2+ based on PET mechanism. More importantly, the stoichiometry of L and Zn2+ was determined to be 2:1 by fluorescent titration, Job's plot method, and ESI-MS spectrometry. The association constant for Zn2+ ions was determined to be 6.26 × 108 M-2, while the limit of detection (LOD) of L was estimated as 5.43 nM, which is a much lower value than WHO and EPA guidelines for drinking water. Moreover, L was successfully applied to detect both Zn2+ and Cu2+ in living cells due to good biocompatibility and excellent low toxicity.