Revealing Gasotransmitter Regulation to Phosphatase Activity in Live Cells by Three‐Channel Imaging Correlation

Enzyme activity in live cells is dynamically regulated by small molecular transmitters for maintaining normal physiological functions. A few probes have been innovated to measure intracellular enzyme activities by fluorescent imaging, but revealing the gasotransmitter regulation to enzyme activity in‐situ remains a long‐standing difficulty. Herein, we report a three‐channel imaging correlation by a single dual‐reactive fluorescent probe to achieve the measurements of phosphatase activity dependence on H2S level in cells. The two reactive sites of probe to H2S and phosphatase individually produce blue and green fluorescent responses, respectively, and the resonance energy transfer can be triggered in the coexistence of them. Fluorescent analysis based on the three‐channel imaging correlation shows that cells have an ideal level of H2S to promote phosphatase activity up to its maximum. Significantly, a slight deviation from the H2S level leads to a sharp decrease of phosphatase activity. The discovery further strengthens our understanding of H2S importance in cellular signalling and various human diseases.

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