Related Experiment Video

Updated: Feb 10, 2026

Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain
09:25

Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain

Published on: May 21, 2019

7.2K

Selective Substrates and Activity-Based Probes for Imaging of the Human Constitutive 20S Proteasome in Cells and

Wioletta Rut1, Marcin Poręba1,2, Paulina Kasperkiewicz1,2

  • 1Department of Bioorganic Chemistry, Faculty of Chemistry , Wroclaw University of Science and Technology , Wyb. Wyspianskiego 27 , 50-370 Wroclaw , Poland.

Journal of Medicinal Chemistry
|May 29, 2018
PubMed

Insights

Researchers developed novel fluorogenic substrates to measure proteasome activity. This method aids in diagnosing diseases linked to altered proteasome function, offering a rapid tool for clinical applications.

Area of Science:

  • Biochemistry and Molecular Biology
  • Enzymology
  • Proteasome Biology

Background:

  • The proteasome is essential for protein homeostasis, and its dysfunction is implicated in diseases like cancer and neurodegeneration.
  • Targeting proteasome activity presents a promising avenue for pharmaceutical development.
  • Existing methods for assessing proteasome activity may lack specificity or simplicity for broad clinical application.

Purpose of the Study:

  • To design and synthesize novel, selective fluorogenic substrates for the three constitutive activities of the 20S proteasome.
  • To develop a rapid and simple method for measuring proteasome activity in biological samples, including whole blood.
  • To enable simultaneous detection of different proteasome subunits using activity-based probes.

Main Methods:

  • Employed the HyCoSuL approach to design and synthesize fluorogenic substrates specific to 20S proteasome activities.
  • Utilized these substrates to assess the inhibitory effects of known proteasome inhibitors (MG-132 and bortezomib).
  • Constructed fluorophore-labeled activity-based probes from selective peptide sequences for simultaneous subunit detection.

Main Results:

  • Successfully designed and synthesized novel fluorogenic substrates demonstrating utility in biochemical assays.
  • Confirmed the effectiveness of the developed substrates in assessing proteasome subunit inhibition by MG-132 and bortezomib.
  • Developed activity-based probes capable of simultaneously detecting constitutive 20S proteasome subunits in cell lysates and red blood cells.

Conclusions:

  • The developed fluorogenic substrates and activity-based probes provide a robust tool for studying proteasome function.
  • The described method offers a simple and rapid approach to measure constitutive 20S proteasome activity in whole human blood.
  • This technique holds potential for the early diagnosis of pathological conditions associated with aberrant proteasome activity.

More Related Videos

Monitoring of Ubiquitin-proteasome Activity in Living Cells Using a Degron dgn-destabilized Green Fluorescent Protein GFP-based Reporter Protein10:25

Monitoring of Ubiquitin-proteasome Activity in Living Cells Using a Degron dgn-destabilized Green Fluorescent Protein GFP-based Reporter Protein

Published on: November 10, 2012

17.2K
Assaying Proteasomal Degradation in a Cell-free System in Plants07:43

Assaying Proteasomal Degradation in a Cell-free System in Plants

Published on: March 26, 2014

15.1K

Related Concept Videos

The Proteasome02:18

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
10.3K
The Proteasome01:13

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
1.7K
The Proteasome02:18

The Proteasome

4.6K
T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...
16.2K
The Proteasome Structure01:17

The Proteasome Structure

The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...
1.7K
Constitutional Isomers of Alkanes02:18

Constitutional Isomers of Alkanes

Organic compounds of the same molecular formula can have different structural formulas called constitutional isomers, and the phenomenon is known as constitutional isomerism. Alkanes with four or more carbons showing multiple structures with the same molecular formula thereby exhibit constitutional isomerism.
The linear isomer of an alkane is prefixed by the term “n”; hence a linear isomer of pentane is known as n-pentane. Based on the type of branching, some of the...
22.8K