Development of Noninvasive Activity-Based Protein Profiling-Bioluminescence Resonance Energy Transfer Platform...

Area of Science:

• Biomedical Research
• Molecular Imaging
• Biochemistry

Background:

• Real-time, longitudinal imaging of protein function in living systems remains a significant challenge.
• High specificity is crucial for understanding complex biological processes.

Purpose of the Study:

• To develop a novel platform for noninvasive, real-time, longitudinal imaging of protein functions.
• To enable the study of protein post-translational modifications in living systems.
• To facilitate target engagement and inhibitor profiling studies in live cells.

Main Methods:

• Platform fusion technology combining activity-based protein profiling (ABPP) with bioluminescence resonance energy transfer (BRET).
• Development of a semisynthetic BRET biosensor.
• Utilizing distance-dependent BRET readout for monitoring protein activity.

Main Results:

• Demonstrated a novel ABPP-BRET method for studying protein function in real time.
• Enabled longitudinal monitoring of post-translational modifications in living systems.
• Successfully applied the method for target engagement and inhibitor profiling in live cells.

Conclusions:

• The ABPP-BRET platform offers a powerful and simple approach for advanced protein activity studies.
• This technology significantly advances the capabilities of activity-based protein profiling.
• Provides unprecedented specificity for imaging protein functions in longitudinal studies.