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Phosphate Ion Targeted Colorimetric and Fluorescent Probe and Its Use to Monitor Endogeneous Phosphate Ion in a Hemichannel-Closed Cell
- Phosphate Ion Targeted Colorimetric and Fluorescent Probe and Its Use to Monitor Endogeneous Phosphate Ion in a Hemichannel-Closed Cell
- Guo, Lin E.; Zhang, Jun Feng; Liu, Xin Yi; Zhang, Li Mei; Zhang, Hong Li; Chen, Jian Hua; Xie, Xiao Guang; Zhou, Ying; Luo, Kaijun; Yoon, Juyoung
- Ewha Authors
- SCOPUS Author ID
- Issue Date
- Journal Title
- ANALYTICAL CHEMISTRY
- 0003-2700; 1520-6882
- vol. 87, no. 2, pp. 1196 - 1201
- AMER CHEMICAL SOC
- SCI; SCIE; SCOPUS
- Fluorescent probe 1, the first inorganic phosphate (Pi) targeted colorimetric and fluorescent probe to detect endogenous Pi in hemichannel-closed cells, has been developed. Probe 1 undergoes a unique Pi induced hydrolytic reaction in DMSOHEPES (V/V = 9:1) buffered (0.02 M, pH 7.4) solutions that produces a colorimetric change associated with a 62 nm red-shift in the UVvis absorption maximum and up to a 780-fold enhancement in the fluorescence intensity. The mechanistic proposal that these spectroscopic changes are associated with reaction Pi with 1 to form coumarin gains support from the results of theoretical calculations and mass spectrometry studies. Observations made in fluorescence imaging studies with HeLa cells and C. elegans show that 1 can be employed to monitor Pi production in vivo caused by apyrase-catalyzed ATP hydrolysis. Moreover, probe 1 was utilized to show that apoptosis of hemichannel-closed Sf9 cells is caused by Inx3 promoted dephosphorylation of Akt (RAC serine/threonine-protein kinase), leading to an elevation of the concentration of Pi. Overall, the study has produced the first fluorescent sensor 1 for endogenous inorganic phosphate. Moreover, the utility of 1 for measuring Pi release in vitro has been demonstrated and utilized to elucidate the mechanism of Inx3 action in hemichannel-closed Sf9 cells.
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