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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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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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Labeling DNA Probes03:31

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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Phosphorylation01:02

Phosphorylation

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The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
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Related Experiment Video

Updated: Mar 31, 2026

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET
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Genetically Encoded Fluorescent Indicators to Visualize Protein Phosphorylation in Living Cells.

Moritoshi Sato1, Yoshio Umezawa2

  • 1Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, 153-8902, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|October 27, 2015
PubMed
Summary

Researchers developed novel fluorescent indicators to visualize protein phosphorylation in living cells. This allows real-time tracking of kinase signaling dynamics, overcoming limitations of traditional methods.

Keywords:
Fluorescence resonance energy transfer (FRET)Fluorescent indicatorsLive cell imagingProtein kinasesProtein phosphorylation reactionsSubcellular dynamic s

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Area of Science:

  • Cellular signaling and molecular biology
  • Biochemistry and biophysics

Background:

  • Protein phosphorylation by intracellular kinases is crucial for cellular signaling pathways.
  • Conventional methods like electrophoresis and immunocytochemistry lack spatial and temporal resolution for studying kinase activity in living cells.
  • Understanding kinase signaling dynamics is essential for deciphering biological processes.

Purpose of the Study:

  • To develop novel tools for visualizing protein phosphorylation in living cells.
  • To overcome the limitations of traditional assays in capturing dynamic kinase signaling.
  • To enable real-time observation of kinase activation and deactivation within single cells.

Main Methods:

  • Development of genetically encoded fluorescent indicators.
  • Application of fluorescence microscopy for live-cell imaging.
  • Visualization of protein phosphorylation and kinase activity in situ.

Main Results:

  • Successful development of fluorescent indicators for protein phosphorylation.
  • Real-time visualization of kinase activation dynamics in single living cells.
  • Spatially and temporally resolved data on kinase signaling pathways.

Conclusions:

  • Genetically encoded fluorescent indicators provide unprecedented insights into kinase signaling.
  • These tools overcome limitations of conventional methods for studying dynamic phosphorylation events.
  • Enables detailed investigation of when, where, and how protein kinases are activated in living cells.