Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

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.

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Understanding attenuated positive facial expression in depressed women: central role of dorsal raphe network alterations and associated serotonin transporter methylation.

Scientific reports·2026
Same author

Enhanced large-scale production and purification of recombinant human NUPR1 for detailed structural and functional insights.

Protein expression and purification·2026
Same author

Revealing hidden noncovalent forces of a disulfide bond in protein interactions and biomolecular condensation.

International journal of biological macromolecules·2026
Same author

Sex differences in pain perception and modulation in the brain: effects of insular cortex stimulation on chronic pain relief.

Brain communications·2025
Same author

NMR investigation of FOXO4-DNA interaction for discriminating target and non-target DNA sequences.

Communications biology·2024
Same author

Mechanism of Methylene Blue Inducing the Disulfide Bond Formation of Tubulin-Associated Unit Proteins.

JACS Au·2024

Related Experiment Video

Updated: Jun 26, 2026

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
10:27

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

Direct characterization of E2-dependent target specificity and processivity using an artificial p27-linker-E2

Kyoung-Seok Ryu1, Yun-Seok Choi, Junsang Ko

  • 1Korea Basic Science Institute, Korea.

BMB Reports
|January 7, 2009
PubMed
Summary

Researchers explored E2 enzyme specificity in ubiquitination. They found specific E2 enzymes, like Cdc34, dictate target protein (p27) modification and ubiquitin linkage types, revealing crucial details about this cellular process.

More Related Videos

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations
10:26

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

Published on: November 7, 2019

In Vitro Analysis of E3 Ubiquitin Ligase Function
06:06

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

Related Experiment Videos

Last Updated: Jun 26, 2026

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
10:27

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta

Published on: December 5, 2019

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations
10:26

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

Published on: November 7, 2019

In Vitro Analysis of E3 Ubiquitin Ligase Function
06:06

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Ubiquitination is a key post-translational modification regulated by E3 ligases and E2 conjugating enzymes.
  • The specificity of E2 enzymes in mediating direct ubiquitination of target proteins remains underexplored, particularly concerning intra-molecular reactions.

Purpose of the Study:

  • To investigate the specificity of E2 enzymes in the ubiquitination of the target protein p27.
  • To determine if E2 enzymes alone can dictate the type of ubiquitin linkage formed.

Main Methods:

  • Construction of artificial E2 fusion proteins tethering the target protein p27 to six different E2 enzymes via a flexible linker.
  • Analysis of the ubiquitination efficiency and linkage types mediated by these fusion proteins.

Main Results:

  • Only three E2 enzymes (UbcH5b, hHR6b, and Cdc34) facilitated intra-molecular ubiquitination of p27.
  • Cdc34 demonstrated high efficiency in subsequent ubiquitination of p27-ubiquitin conjugates.
  • Specific residues (184-196) in Cdc34 were identified as crucial for possessive ubiquitination of p27.
  • E2 enzyme identity alone determined the differential ubiquitin linkage formation.

Conclusions:

  • Direct E2 enzyme specificity for the target protein p27 was demonstrated.
  • E2 enzymes play a critical role in determining both the site and type of ubiquitination.
  • These findings provide new insights into the regulatory mechanisms of ubiquitination and its impact on cellular processes.