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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: Aug 28, 2025

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Published on: November 28, 2018

Development and Characterization of Flavin-Binding Fluorescent Proteins, Part I: Basic Characterization.

Alina Remeeva¹, Anna Yudenko¹, Vera V Nazarenko¹

¹Research Center for Molecular Mechanisms of Aging and Age-Related Diseases, Moscow Institute of Physics and Technology, Dolgoprudny, Russia.

Methods in Molecular Biology (Clifton, N.J.)

|September 15, 2022

Summary

This summary is machine-generated.

Flavin-based fluorescent proteins (FbFPs) are versatile reporters derived from light-oxygen-voltage (LOV) domains. This work details methods for identifying, designing, and characterizing these useful proteins for in vivo imaging.

Keywords:

Flavin Flavin-binding fluorescent protein Fluorescence microscopy Genome mining LOV domain

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Area of Science:

Biochemistry
Molecular Biology
Biophysics

Background:

Flavin-based fluorescent proteins (FbFPs) are derived from light-oxygen-voltage (LOV) domains.
They utilize endogenous flavins as chromophores.
FbFPs function as versatile in vivo reporter proteins under aerobic and anaerobic conditions.

Purpose of the Study:

To present a comprehensive methodology for working with FbFPs.
To guide the identification and design of novel FbFPs.
To enable basic fluorescence microscopy experiments using FbFPs.

Main Methods:

Genomic database searches for LOV domain sequences.
Rational design of new FbFP constructs.
Biochemical, spectroscopic, photophysical, and photochemical characterization.
Basic fluorescence microscopy.

Main Results:

Established protocols for FbFP identification and design.
Characterized key properties of FbFPs.
Demonstrated utility in fluorescence microscopy.

Conclusions:

FbFPs are adaptable tools for in vivo protein research.
The presented methods facilitate the development and application of FbFPs.
FbFPs offer a robust reporter system for diverse biological applications.