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Related Experiment Video

Updated: Jan 20, 2026

Monitoring of Ubiquitin-proteasome Activity in Living Cells Using a Degron dgn-destabilized Green Fluorescent Protein GFP-based Reporter Protein
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Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes.

Sabine Schipper-Krom1, Alicia Sanz Sanz1, Emma J van Bodegraven1

  • 1Department of Medical Biology, Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands.

Frontiers in Molecular Biosciences
|September 5, 2019
PubMed
Summary

This study compares methods for studying proteasome function in living cells, crucial for understanding protein degradation and disease. Techniques include fluorescent tags and activity probes to analyze proteasome localization, dynamics, and activity.

Keywords:
activity probesdynamicsfluorescenceliving cellsproteasome

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • The proteasome is a vital molecular machine responsible for protein degradation in eukaryotic cells.
  • Dysfunctional proteasomes are implicated in various human diseases.
  • Understanding proteasome activity is critical for disease research.

Purpose of the Study:

  • To describe and compare methods for studying proteasome function in living cells.
  • To provide tools for investigating proteasome localization, dynamics, and activity.

Main Methods:

  • Utilizing fluorescently tagged proteasome subunits for visualization.
  • Employing activity-based proteasome probes (ABPs) for activity assessment.
  • Integrating tagged proteasomes and ABPs in biochemical and microscopy assays.

Main Results:

  • Demonstrated utility of fluorescent tags for tracking proteasome movement and localization.
  • Validated ABPs for quantifying proteasome activity in vitro and in situ.
  • Showcased combined approaches for comprehensive proteasome analysis.

Conclusions:

  • Fluorescent tags and activity probes offer powerful complementary methods for studying proteasome function.
  • These techniques enable detailed investigation of proteasome dynamics and activity in living cells.
  • Advancing proteasome research is key to understanding and treating associated diseases.