Following a protein kinase activity using a field-effect transistor device.

Area of Science:

• Biochemistry
• Biosensor Technology
• Molecular Biology

Background:

• Protein kinases play crucial roles in cellular signaling pathways.
• Monitoring protein kinase activity is essential for understanding cellular processes and disease mechanisms.
• Existing methods for kinase activity detection can be complex and time-consuming.

Purpose of the Study:

• To develop a novel electronic biosensor for real-time monitoring of casein kinase II activity.
• To investigate the feasibility of regenerating the sensor surface for repeated measurements.
• To enable electronic readout of enzymatic activity using a peptide-functionalized ion-sensitive field-effect transistor.

Main Methods:

• Fabrication of a peptide-functionalized ion-sensitive field-effect transistor (ISFET) biosensor.
• Phosphorylation of the peptide surface by casein kinase II in the presence of ATP.
• Electronic measurement of the ISFET signal to quantify kinase activity.
• Regeneration of the sensor surface using alkaline phosphatase.

Main Results:

• The ISFET biosensor successfully detected casein kinase II activity through specific phosphorylation events.
• The electronic signal directly correlated with the enzymatic activity.
• Treatment with alkaline phosphatase effectively regenerated the active sensing surface, allowing for multiple uses.
• The developed system provides a sensitive and reusable platform for kinase activity detection.

Conclusions:

• Peptide-functionalized ISFETs offer a promising platform for electronic detection of protein kinase activity.
• The sensor exhibits reusability after regeneration, enhancing its practical applicability.
• This technology has potential applications in biochemical research and diagnostics.