A water-soluble fluorescent probe for the sensitive detection of endogenous alkaline phosphatase in living cells

作者:Xu, Longbin; He, Xu; Ma, Pinyi*; Liu, Xin; Zhang, Fangmei; Sun, Ying; Song, Daqian; Wang, Xinghua*
来源:Dyes and Pigments, 2018, 159: 584-589.
DOI:10.1016/j.dyepig.2018.07.023

摘要

Alkaline phosphatase (ALP) is an essential enzyme in various tissues, which has been reported to be extensively involved in various physiological and pathological processes. Fluorescent probe assays carried out in vivo are desirable for monitoring the ALP activity. In this study, a novel red-light fluorescent DHP probe, which exhibited good selectivity and high sensitivity, for monitoring the ALP activity in real time was described. The DHP probe exhibited good water solubility, and the detection limit of ALP was determined as 0.031 U/L; this value is almost less than those of the other probes reported previously. The probe exhibited high selectivity, which will not interfere with various enzymes and amino acids. Moreover, the sensing and luminescence mechanisms of ALP to DHP was investigated by density functional theory (DFT) and time-dependent DFT. Furthermore, the DHP probe was applied for the detection of endogenous ALP in BEL-7402 cells, and the fluctuation of the ALP levels in BEL7402 and HEK 293 T cells was successfully tracked by fluorescence microscopy.