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Related Concept Videos

Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.

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Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations
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Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Published on: November 7, 2019

Efficient approaches for characterizing ubiquitinated proteins.

Roland Hjerpe1, Manuel S Rodríguez

  • 1Ubiquitin-Like Molecules and Cancer Laboratory, Proteomics Unit, CIC bioGUNE (Asociacion Centro de Investigación Cooperativa en Biociencias), CIBERehd, Bizkaia Technology Park, Building 801A, 48160 Derio, Spain.

Biochemical Society Transactions
|September 17, 2008
PubMed
Summary

Detecting ubiquitinated proteins is challenging due to rapid deubiquitination by deubiquitinating enzymes (DUBs) and proteasomal degradation. This review covers practical methods for detecting and purifying ubiquitinated proteins, focusing on ubiquitin-binding domain affinity purification.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Post-translational modifications rapidly alter protein activity within cells.
  • Ubiquitination, the covalent attachment of ubiquitin, significantly impacts protein biochemical properties.
  • Detecting ubiquitinated proteins is difficult due to their labile nature and rapid reversal by deubiquitinating enzymes (DUBs) and proteasomal degradation.

Purpose of the Study:

  • To review practical approaches for the detection, purification, and characterization of ubiquitinated proteins.
  • To highlight challenges in identifying ubiquitinated protein species.
  • To focus on the development and application of ubiquitin-binding domain (UBD)-ubiquitin affinity purification.

Main Methods:

  • Review of established and emerging techniques for ubiquitinated protein detection.
  • Appraisal of histidine-tagged ubiquitin methods.
  • Detailed examination of ubiquitin-binding domain (UBD)-ubiquitin affinity purification strategies.

Main Results:

  • Ubiquitination is a critical but transient post-translational modification.
  • Deubiquitinating enzymes (DUBs) and proteasomal degradation complicate the detection of ubiquitinated proteins.
  • UBD-ubiquitin affinity purification offers a robust method for isolating and studying ubiquitinated proteins.

Conclusions:

  • Effective detection and purification of ubiquitinated proteins are essential for understanding their biological roles.
  • Challenges in detecting labile ubiquitinated species necessitate advanced methodologies.
  • UBD-ubiquitin affinity purification represents a significant advancement in the study of ubiquitination.