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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...

Protein Diffusion in the Membrane

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Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...

Overview of Protein Sorting and Transport

Overview of Protein Sorting and Transport

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Related Experiment Video

Updated: Mar 19, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Global, quantitative and dynamic mapping of protein subcellular localization.

Daniel N Itzhak¹, Stefka Tyanova¹, Jürgen Cox¹

¹Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.

|June 10, 2016

Summary

Dynamic Organellar Maps provide a proteomic method to globally map protein movements within cells. This technique offers a quantitative view of cell anatomy and organellar composition, revealing protein translocation events without specific reagents.

Keywords:

EGF signalling cell biology computational biology human map of the cell organellar proteomics protein translocations spatial proteomics subcellular localization systems biology

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Related Experiment Videos

Last Updated: Mar 19, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

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Published on: June 30, 2018

Enriching Subcellular Proteins in Leptospira Using a Triton X-114-Based Fractionation Approach

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Heterokaryon Technique for Analysis of Cell Type-specific Localization

Heterokaryon Technique for Analysis of Cell Type-specific Localization

Published on: March 11, 2011

Area of Science:

Cell Biology
Proteomics
Systems Biology

Background:

Subcellular localization is crucial for protein function and cellular regulation.
Existing methods lack the ability to globally map protein translocation events.

Purpose of the Study:

To develop and apply Dynamic Organellar Maps, a novel proteomic method for global mapping of protein translocation.
To generate a comprehensive database of protein localization and copy number in HeLa cells.
To quantitatively model dynamic protein movements in response to stimuli.

Main Methods:

Development and application of Dynamic Organellar Maps (proteomic method).
Static mapping to create a database of protein localization and absolute copy number for over 8700 HeLa cell proteins.
Dynamic mapping to capture protein translocation events after EGF stimulation.

Main Results:

Generated a database with localization and copy number for >8700 HeLa cell proteins, covering major organelles with >92% accuracy.
Provided a quantitative view of HeLa cell anatomy and organellar composition at the protein level.
Captured and modeled dynamic protein translocation events following EGF stimulation.

Conclusions:

Dynamic Organellar Maps offer a powerful tool for proteome-wide analysis of physiological protein movements.
The method is reagent-independent, ensuring broad applicability across cell biology research.
This approach enables a deeper understanding of cellular regulation through protein localization dynamics.