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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Published on: July 25, 2014

Relocation sensors to quantify signaling dynamics in live single cells.

Victoria Wosika¹, Serge Pelet¹

¹Department of Fundamental Microbiology, University of Lausanne, 1015 Lausanne, Switzerland.

Current Opinion in Biotechnology

|January 29, 2017

Summary

Cellular heterogeneity impacts responses to stimuli. Fluorescent biosensors, particularly relocation reporters, enable real-time monitoring of single-cell signal transduction dynamics for kinase activity and transcription factor activation.

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Published on: March 18, 2021

Area of Science:

Cell Biology
Biophysics
Molecular Biology

Background:

Cellular heterogeneity in shape, cell-cycle stage, and protein expression influences cellular responses.
Understanding these variations is crucial for elucidating signal transduction pathways.
Fluorescence microscopy offers a powerful tool for visualizing cellular differences and real-time responses.

Purpose of the Study:

To highlight the importance of cellular heterogeneity in modulating cell responses.
To introduce fluorescent biosensors as tools for studying live single-cell dynamics.
To emphasize the role of relocation reporters in quantifying signal transduction.

Main Methods:

Utilizing fluorescence microscopy to visualize cellular heterogeneity and dynamics.
Developing and employing fluorescent biosensors for live single-cell analysis.
Implementing relocation reporters to monitor signal transduction pathways.

Main Results:

Demonstrated that cellular variations impact responses to environmental stimuli.
Showcased the utility of fluorescent biosensors in observing key cellular events in real-time.
Validated relocation reporters for quantifying signal transduction dynamics.

Conclusions:

Cellular heterogeneity is a key factor in biological responses.
Fluorescent biosensors, especially relocation reporters, are effective for live single-cell analysis of signal transduction.
These methods provide insights into kinase activity, transcription factor activation, and protein expression.