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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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Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Published on: October 21, 2016

Extrinsic fluorescent dyes as tools for protein characterization.

Andrea Hawe¹, Marc Sutter, Wim Jiskoot

¹Division of Drug Delivery Technology, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands.

Pharmaceutical Research

|January 4, 2008

Summary

Extrinsic fluorescent dyes are versatile tools for protein analysis, aiding in the study of folding, hydrophobicity, and aggregation. This review covers their mechanisms, spectral properties, and diverse applications in characterizing proteins.

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Area of Science:

Biochemistry and Biophysics
Protein Science
Fluorescence Spectroscopy

Background:

Noncovalent, extrinsic fluorescent dyes are crucial for protein analysis, including characterizing folding intermediates, measuring surface hydrophobicity, and detecting protein aggregation or fibrillation.
Mechanisms like solvent relaxation and intramolecular charge transfer govern dye fluorescence, influenced by environmental factors and protein interactions.

Observation:

The use of extrinsic fluorescent dyes such as 8-anilino-1-naphthalenesulfonic acid (ANS), Bis-ANS, Nile Red, and Thioflavin T has significantly increased.
These dyes offer versatility, high sensitivity, and suitability for high-throughput screening in protein research.

Findings:

This review provides a comprehensive overview of available extrinsic fluorescent dyes.
It details their spectral properties and presents illustrative examples of their applications in protein characterization.

Implications:

Understanding these dyes enhances protein analysis techniques.
Their application facilitates advancements in drug discovery and understanding protein misfolding diseases.