Sensitive fluorescent biosensor based on a europium-based metal-organic framework for protein kinase activity...

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biomedical Engineering

Background:

• Protein kinases are vital regulators of cellular processes, making their activity crucial for human health.
• Dysregulated protein kinase activity is implicated in various human diseases, necessitating accurate monitoring methods.
• Existing analytical techniques for protein kinase activity present limitations, driving the need for innovative approaches.

Purpose of the Study:

• To develop a novel fluorescent biosensor for the sensitive and selective detection of protein kinase activity.
• To utilize the Eu(BTC) (H2O)6 metal-organic framework as the core component of the biosensor.
• To evaluate the biosensor's performance in detecting kinase activity in cellular environments and screening for inhibitors.

Main Methods:

• Development of a fluorescent biosensor based on Eu(BTC) (H2O)6.
• Utilizing a fluorophore-labeled substrate polypeptide that is phosphorylated by protein kinase.
• Coordination of the phosphorylated peptide with the europium site of Eu(BTC) (H2O)6 for fluorescence detection.

Main Results:

• The biosensor demonstrated a low detection limit of 0.00003 U μL-1 with excellent selectivity and universality.
• Successfully detected protein kinase activity within MCF-7 cells.
• Identified H-89 and ellagic acid as inhibitors of protein kinase activity with IC50 values of 1.09 and 19.88 nmol L-1, respectively.

Conclusions:

• The developed Eu(BTC) (H2O)6-based fluorescent biosensor is a sensitive and versatile tool for monitoring protein kinase activity.
• This biosensor shows significant potential for applications in disease diagnostics and the screening of kinase inhibitors.
• The method provides a valuable advancement for understanding and controlling protein kinase functions in biological systems.