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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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Visualizing Low-Abundance Proteins and Post-Translational Modifications in Living Drosophila Embryos via Fluorescent Antibody Injection

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Real-time imaging of protein internalization using aptamer conjugates.

Wei Li¹, Xiaohai Yang, Kemin Wang

¹College of Chemistry and Chemical Engineering, Engineering Center for Biomedicine, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, PR China.

Analytical Chemistry

|June 7, 2008

Summary

Researchers visualized how angiogenin, a tumor angiogenesis factor, enters cells using a fluorescent aptamer probe. This novel method tracks protein internalization in real-time within endothelial and cancer cells.

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Published on: October 10, 2010

Area of Science:

Biochemistry
Cell Biology
Molecular Medicine

Background:

Angiogenin is a key factor in tumor angiogenesis.
Understanding protein internalization is crucial for targeted therapies.

Purpose of the Study:

To develop a method for visualizing angiogenin cellular internalization.
To investigate the real-time uptake of aptamer-angiogenin conjugates.

Main Methods:

Utilized fluorophore-labeled aptamers conjugated with angiogenin.
Employed confocal laser scanning microscopy for real-time imaging.
Performed nuclear staining and Z-axis scanning for intracellular localization.

Main Results:

Aptamer-angiogenin conjugates selectively bound to human umbilical vein endothelial cells (HUVECs) and MCF-7 breast cancer cells.
Conjugates were internalized into intracellular organelles.
Dynamic imaging confirmed rapid internalization of the conjugates.

Conclusions:

A novel, simple method was established to visualize protein internalization in real-time.
Fluorophore-labeled aptamers serve as effective fluorescent probes for tracking protein uptake.
This technique offers insights into the spatiotemporal dynamics of protein internalization.