The first fluorogenic sensor for sphingosine-1-phosphate lyase activity in intact cells

Area of Science:

• Biochemistry
• Cell Biology
• Analytical Chemistry

Background:

• Sphingosine-1-phosphate lyase (S1PL) is a key enzyme in sphingolipid metabolism.
• Accurate measurement of S1PL activity is crucial for understanding cellular processes and disease states.
• Existing methods for S1PL activity determination may have limitations in sensitivity or applicability.

Purpose of the Study:

• To develop novel fluorogenic sensors for quantifying sphingosine-1-phosphate lyase (S1PL) activity.
• To optimize sensors for suitable kinetic parameters and high sensitivity.
• To enable S1PL activity measurement in both cell lysates and intact cells.

Main Methods:

• Development of new fluorogenic sensor molecules.
• Characterization of sensor kinetic parameters and sensitivity.
• Loading of the RBM148 probe into cationic liposomes.
• Determination of S1PL activity in cell lysates using the developed sensors.
• Assessment of S1PL activity in intact cells using the RBM148-liposome probe.

Main Results:

• Novel fluorogenic sensors with appropriate kinetic properties and sensitivity were successfully synthesized.
• The RBM148 probe demonstrated efficient loading into cationic liposomes.
• The RBM148-liposome system effectively determined sphingosine-1-phosphate lyase activity in cell lysates.
• The RBM148 probe was successfully utilized to measure S1PL activity in intact cells.

Conclusions:

• New fluorogenic sensors provide a sensitive and efficient method for determining sphingosine-1-phosphate lyase activity.
• The RBM148 probe, particularly when encapsulated in liposomes, offers a versatile tool for S1PL activity analysis in various biological contexts.
• These advancements facilitate further research into the role of S1PL in cellular functions and disease.