Can we see PIP(3) and hydrogen peroxide with a single probe?

Area of Science:

• Cell Biology
• Biochemistry
• Molecular Imaging

Background:

• Phosphatidylinositol 3-kinase (PI3K) signaling is crucial in cell growth and survival.
• Hydrogen peroxide (H2O2) is a key signaling molecule in cellular processes.
• Simultaneous measurement of these pathways is challenging.

Purpose of the Study:

• To develop a genetically encoded sensor for parallel measurement of PI3K activity and H2O2 levels.
• To monitor the dynamics of these molecules in living cells.
• To establish a prototype for multifunctional fluorescent sensors.

Main Methods:

• Development of a genetically encoded sensor (PIP-SHOW).
• Utilized sensor translocation to plasma membrane for PIP3 detection.
• Employed ratiometric excitation changes for simultaneous H2O2 detection.
• Applied sensor in platelet-derived growth factor-stimulated fibroblasts and T-lymphocytes.

Main Results:

• PIP-SHOW successfully translocates upon PI3K pathway activation.
• The sensor enables rapid, simultaneous reporting of H2O2 concentration changes.
• Dynamics of PIP3 and H2O2 were monitored in stimulated fibroblasts and T-cells.
• Demonstrated the sensor's utility in complex cellular environments like immunological synapses.

Conclusions:

• PIP-SHOW provides a novel method for parallel monitoring of PI3K activity and H2O2.
• The sensor facilitates real-time analysis of cellular signaling crosstalk.
• PIP-SHOW serves as a valuable prototype for developing advanced, multi-analyte fluorescent sensors.