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Related Experiment Video

Updated: Feb 5, 2026

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

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Published on: January 27, 2012

Visualizing PINK1 Activity Dynamics in Single Cells with a Phase Separation-Based Kinase Activity Reporter.

Katie G Vineall¹, Alexia Andrikopoulos¹, Michael J Sun¹

¹Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, California 90095, United States.

|February 3, 2026

Summary

Researchers developed a novel biosensor, PINK1-SPARK, to track the activity of Phosphatase and tensin homologue-induced kinase 1 (PINK1) in real-time within single cells. This tool illuminates PINK1 dynamics and heterogeneity, advancing studies on mitophagy and cellular energy regulation.

Keywords:

PINK1 biosensor fluorescence kinase activity reporter mitophagy

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Last Updated: Feb 5, 2026

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Area of Science:

Cell Biology
Biochemistry
Molecular Biology

Background:

Phosphatase and tensin homologue-induced kinase 1 (PINK1) is crucial for mitophagy, cell death, and cellular bioenergetics.
Existing bulk methods for studying PINK1 activity provide limited temporal resolution and miss cell-to-cell variations.

Purpose of the Study:

To develop a novel biosensor for real-time monitoring of PINK1 kinase activity in single live cells.
To overcome limitations of bulk assays and characterize PINK1 activity dynamics and heterogeneity.

Main Methods:

Developed a phase separation-based PINK1 biosensor (PINK1-SPARK) utilizing the SPARK design.
Utilized PINK1-SPARK to observe real-time PINK1 activity in response to mitochondrial depolarization and pharmacological activation.
Created a HaloTag-based PINK1-SPARK for multiplexed imaging with mitochondrial damage markers.

Main Results:

Successfully visualized real-time PINK1 kinase activity in single cells.
Demonstrated the ability to track dynamic changes in PINK1 activity under various cellular conditions.
Enabled simultaneous imaging of PINK1 activity and mitochondrial damage markers.

Conclusions:

PINK1-SPARK is a novel tool for temporal measurement of PINK1 activity in single live cells.
This biosensor facilitates deeper understanding of PINK1's role in mitophagy and cellular function.
The tool allows for detailed characterization of cell-to-cell heterogeneity in PINK1 activity.