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Updated: Apr 22, 2026
BRET-based G Protein Biosensors for Measuring G Protein-Coupled Receptor Activity in Live Cells
Published on: November 7, 2025
Assessing GPCR and G protein signaling to the nucleus in live cells using fluorescent biosensors
1Department of Pharmacology, University of California, Davis, CA, 95616-8636, USA.
Abstract:
G protein-coupled receptor (GPCR) signaling cascades regulate a wide variety of cellular processes and feature prominently in many cardiovascular pathologies. As such they represent major drug targets and discovering novel aspects of GPCR signaling provide important opportunities to identify additional potential therapeutic approaches to reverse or prevent cardiac remodeling and failure. Monitoring cellular trafficking of signaling components and specific protein kinase activities using fluorescent biosensors has provided key insight into stress/GPCR-induced kinase signaling networks and their effect on cardiac gene expression. Herein we describe the protocols for the expression, visualization (by confocal microscopy), and interpretation of data obtained with such biosensors expressed in adult cardiomyocytes. Our focus is on the cellular trafficking of class II histone deacetylases (i.e., HDAC5) and on the FRET sensor (Camui) for calmodulin-dependent protein kinase II (CaMKII).
Insights
This study details methods for visualizing G protein-coupled receptor (GPCR) signaling in heart cells using fluorescent biosensors. These techniques help understand how protein movement and kinase activity impact cardiac health and disease.
Area of Science:
- Cardiovascular Biology
- Molecular Cell Biology
- Biochemistry
Background:
- G protein-coupled receptor (GPCR) signaling is crucial in cellular processes and cardiovascular diseases, making them key drug targets.
- Understanding GPCR signaling networks offers opportunities for novel therapeutic strategies against cardiac remodeling and failure.
Purpose of the Study:
- To describe protocols for expressing and visualizing fluorescent biosensors in adult cardiomyocytes.
- To provide methods for interpreting data on cellular trafficking and kinase activity related to GPCR signaling.
Main Methods:
- Utilizing fluorescent biosensors to monitor cellular trafficking of signaling components, specifically class II histone deacetylases (HDAC5).
- Employing a Förster Resonance Energy Transfer (FRET) sensor (Camui) to detect calmodulin-dependent protein kinase II (CaMKII) activity.
- Confocal microscopy for visualization and data interpretation.
Main Results:
- Established protocols for biosensor expression and imaging in cardiomyocytes.
- Demonstrated the utility of biosensors in studying stress/GPCR-induced kinase signaling networks.
- Provided insights into the cellular trafficking of HDAC5 and CaMKII activity.
Conclusions:
- Fluorescent biosensors are powerful tools for dissecting GPCR signaling pathways in the heart.
- These methods facilitate the study of kinase signaling networks and their impact on cardiac gene expression.
- The described protocols can advance the discovery of new therapeutic targets for cardiovascular pathologies.
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