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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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Biofunctionalized Prussian Blue Nanoparticles for Multimodal Molecular Imaging Applications

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Fluorescent, bioactive protein nanoparticles (prodots) for rapid, improved cellular uptake.

Inoka K Deshapriya¹, Bobbi S Stromer¹, Ajith Pattammattel¹

¹†Department of Chemistry and Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut 06269-3060, United States.

Bioconjugate Chemistry

|February 3, 2015

Summary

Researchers developed fluorescent protein nanoparticles (Prodots) for rapid cellular uptake. These bioactive nanoparticles, made from proteins like bovine serum albumin and enzymes, show potential for cellular imaging and controlled delivery.

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Area of Science:

Biotechnology
Nanotechnology
Cell Biology

Background:

Developing novel nanoparticles for cellular applications is crucial.
Protein-based nanoparticles offer biocompatibility and biodegradability advantages.
Efficient cellular uptake mechanisms are needed for targeted delivery and imaging.

Purpose of the Study:

To synthesize highly fluorescent, bioactive protein-based nanoparticles (Prodots).
To investigate the facile uptake of these Prodots into the cytoplasm of cells.
To explore chemical triggers that control the cellular uptake process.

Main Methods:

Synthesis of Prodots from various proteins (e.g., bovine serum albumin, enzymes).
Characterization using gel electrophoresis, TEM, CD, fluorescence spectroscopy, DLS.
Cellular uptake studies using optical microscopy and controlled chemical triggers.

Main Results:

Prodots (15-50 nm) retained significant protein secondary structure and enzyme activity.
Prodots exhibited enhanced stability with increased denaturation temperatures and storage half-lives.
Rapid cellular uptake (1-3 h) was observed, facilitated by glucose oxidase and glucose, and induced by hydrogen peroxide.

Conclusions:

A simple method for creating bioactive, fluorescent protein nanoparticles was established.
Cellular uptake of Prodots is rapid and controllable via chemical triggers.
These Prodots are promising for cellular imaging and drug delivery applications.