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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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Related Experiment Video

Updated: Dec 25, 2025

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Temperature sensing using red fluorescent protein.

Kanagavel Deepankumar¹, Saravanan Prabhu Nadarajan², Dong-Ho Bae²

¹1School of Biotechnology, Yeungnam University, Gyeongsan, 712-749 Korea.

Biotechnology and Bioprocess Engineering : BBE

|March 29, 2020

Summary

This summary is machine-generated.

Genetically encoded fluorescent proteins like mRFP1 can measure intracellular temperature. This study shows mRFP1 and its variants are feasible as in vivo temperature sensors.

Keywords:

4-fluoroproline mRFP1 non-canonical amino acid incorporation temperature sensor thermal stability

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Area of Science:

Biophysics
Molecular Biology
Cell Biology

Background:

Genetically encoded fluorescent proteins are crucial for biological imaging.
Accurate measurement of intracellular temperature is vital for understanding cellular processes.

Purpose of the Study:

To investigate the potential of mRFP1 and its variants as intracellular thermometers.
To quantify the temperature sensitivity of these fluorescent proteins.

Main Methods:

Expression of mRFP1 and its variants in E. coli.
Measurement of fluorescence intensity at varying temperatures.
Calculation of temperature sensitivity.

Main Results:

A linear correlation was found between temperature and fluorescence intensity for mRFP1 and its variants.
Temperature sensitivities were determined as -1.27%/°C for mRFP1, -1.26%/°C for mRFP-P63A, and -0.77%/°C for mRFP-P63A[(4R)-FP].

Conclusions:

mRFP1 and its variants demonstrate feasibility as effective in vivo temperature sensors.
These fluorescent proteins offer a novel tool for monitoring cellular thermal dynamics.