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.
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Pinching-off of Coated Vesicles01:32

Pinching-off of Coated Vesicles

Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
Protein Folding01:22

Protein Folding

Overview
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
Nuclear Protein Sorting01:34

Nuclear Protein Sorting

Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...

You might also read

Related Articles

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

Sort by
Same author

The E3-ome gene-centric compendium reveals the human E3 ligase landscape.

Cell·2026
Same author

Integrase anchors viral RNA to the HIV-1 capsid interior.

Nature·2026
Same author

Structure and mechanism of the HECT ligase HECTD3.

Nature communications·2026
Same author

Capsid flexibility during Ty1 virus-like particle assembly.

bioRxiv : the preprint server for biology·2025
Same author

KHNYN is a manganese-dependent endoribonuclease required for ZAP-mediated antiviral restriction.

Nucleic acids research·2025
Same author

Covalent fragment screening to inhibit the E3 ligase activity of bacterial NEL enzymes SspH1 and SspH2.

RSC chemical biology·2025

Related Experiment Video

Updated: Jun 23, 2026

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
13:02

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO

Published on: December 28, 2019

NEMO oligomerization and its ubiquitin-binding properties.

Frank J Ivins1, Mark G Montgomery, Susan J M Smith

  • 1Division of Molecular Structure, MRC-National Institute for Medical Research, The Ridgeway, London NW71AA, UK.

The Biochemical Journal
|May 9, 2009
PubMed
Summary

The NF-kappaB essential modulator (NEMO) protein forms dimers and tetramers, with tetramerization disrupted by IKKbeta interaction. NEMO binds di-ubiquitin, suggesting a threshold mechanism for NF-kappaB pathway activation.

More Related Videos

In Vitro Analysis of E3 Ubiquitin Ligase Function
06:06

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
11:36

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones

Published on: July 25, 2019

Related Experiment Videos

Last Updated: Jun 23, 2026

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO
13:02

Production, Crystallization, and Structure Determination of the IKK-binding Domain of NEMO

Published on: December 28, 2019

In Vitro Analysis of E3 Ubiquitin Ligase Function
06:06

In Vitro Analysis of E3 Ubiquitin Ligase Function

Published on: May 14, 2021

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
11:36

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones

Published on: July 25, 2019

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Protein Structure

Background:

  • The IKK complex regulates NF-kappaB activation by degrading IkappaB.
  • NEMO, the IKK complex's regulatory subunit, integrates upstream signals, particularly polyubiquitin chains.
  • The oligomeric state of NEMO and its activation mechanism remain unclear.

Purpose of the Study:

  • To investigate the oligomeric state of NEMO.
  • To elucidate the mechanism of IKK complex activation by NEMO.
  • To understand NEMO's interaction with ubiquitin chains and IKK subunits.

Main Methods:

  • Hydrodynamic techniques were employed to study NEMO's oligomeric state.
  • Interactions between NEMO, IKKbeta peptides, and di-ubiquitin were analyzed.
  • Stoichiometry of NEMO-di-ubiquitin binding was determined using various NEMO constructs.

Main Results:

  • Apo-NEMO exists as an elongated dimer in equilibrium with a tetramer.
  • IKKbeta peptide interaction prevents NEMO tetramerization, indicating mutual exclusivity.
  • NEMO binds linear di-ubiquitin with a 1:1 dimer stoichiometry, with a second weaker site appearing at higher concentrations.

Conclusions:

  • NEMO's oligomerization state is dynamic and regulated by interactions with IKK subunits.
  • Two distinct NEMO-di-ubiquitin complexes are formed, potentially acting as an activation threshold.
  • This mechanism ensures specificity in NF-kappaB signaling.