Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Immunoglobulin-like Cell Adhesion Molecules

Immunoglobulin-like Cell Adhesion Molecules

Immunoglobulin-like cell adhesion molecules or Ig-CAMs are a versatile group of cell surface glycoproteins belonging to the immunoglobulin protein superfamily. Ig-CAMs possess the characteristic immunoglobulin protein domains and other domains such as the fibronectin type III domain. The Ig domains are glycosylated to varying degrees in different Ig-CAMs.
Ig-CAMs exhibit either homophilic binding (to other Ig-CAMs) or heterophilic binding (to other ligands such as integrins). While most Ig-CAMs...

NF-κB-dependent Signaling Pathway

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...

Anchoring Junctions

Anchoring Junctions

Anchoring junctions are multiprotein complexes that help cells connect to other cells and the extracellular matrix. Anchoring junctions are present on the lateral and basal surfaces of cells, providing strong and flexible connections. Focal adhesions are often formed due to cell interactions with the ECM substrata, which initiate signal transduction via kinase cascades and other mechanisms. Together, they provide stability and tissue integrity. There are three types of anchoring junctions:...

NF-kB-dependent Signaling Pathway

NF-kB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...

You might also read

Related Articles

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

Sort by

Same author

Development of a Model Protein Interaction Pair as a Benchmarking Tool for the Quantitative Analysis of 2-Site Protein-Protein Interactions.

Journal of biomolecular techniques : JBT·2015

Same author

Structural and functional analysis of betaine aldehyde dehydrogenase from Staphylococcus aureus.

Acta crystallographica. Section D, Biological crystallography·2015

Same author

Why you should be using more SPR biosensor technology.

Drug discovery today. Technologies·2014

Same author

A bacteriophage capsid protein provides a general amyloid interaction motif (GAIM) that binds and remodels misfolded protein assemblies.

Journal of molecular biology·2014

Same author

Biophysical fragment screening of the β1-adrenergic receptor: identification of high affinity arylpiperazine leads using structure-based drug design.

Journal of medicinal chemistry·2013

Same author

CCR5 is a receptor for Staphylococcus aureus leukotoxin ED.

Nature·2012

Search research articles

Related Experiment Video

Updated: Jul 7, 2026

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

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

Published on: December 28, 2019

High-affinity interaction between IKKbeta and NEMO.

Yu-Chih Lo¹, Upendra Maddineni, Jee Y Chung

¹Department of Biochemistry, Weill Medical College of Cornell University, New York, New York 10021, USA.

|February 13, 2008

Summary

This summary is machine-generated.

The NEMO-binding domain peptide alone is insufficient for IKKbeta interaction. A longer C-terminal region of IKKbeta binds NEMO with high affinity, forming a stable complex.

More Related Videos

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

Published on: November 2, 2018

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Published on: April 21, 2015

Related Experiment Videos

Last Updated: Jul 7, 2026

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

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

Published on: December 28, 2019

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

Published on: November 2, 2018

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Co-immunoprecipitation of the Mouse Mx1 Protein with the Influenza A Virus Nucleoprotein

Published on: April 21, 2015

Area of Science:

Molecular Biology
Biochemistry
Cell Signaling

Background:

The inhibitor of kappaB kinase (IKK) complex, including IKKalpha/IKKbeta and NEMO, is crucial for nuclear factor kappaB (NF-kappaB) pathway activation.
NF-kappaB signaling is a key regulator of immune responses, inflammation, and cell survival.
The NEMO-binding domain (NBD) in IKKalpha/IKKbeta is known to mediate interaction with NEMO.

Purpose of the Study:

To investigate the binding affinity and stoichiometry of the interaction between NEMO and the C-terminal region of IKKbeta.
To determine if the previously identified NBD is sufficient for high-affinity NEMO binding.
To characterize the biophysical properties of the NEMO-IKKbeta C-terminal interaction.

Main Methods:

Surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) for quantitative binding measurements.
Multiangle light scattering (MALS) coupled with refractive index detection for complex stoichiometry analysis.
Biochemical characterization of protein-protein interactions.

Main Results:

The NEMO-binding domain (NBD) peptide of IKKbeta alone does not mediate high-affinity binding to NEMO.
A longer C-terminal region of IKKbeta exhibits high affinity for NEMO.
SPR and ITC confirmed differential binding affinities and provided kinetic and thermodynamic insights.
MALS analysis revealed that the longer IKKbeta C-terminal region forms a stable 2:2 stoichiometric complex with NEMO.

Conclusions:

The NEMO-binding site in IKKbeta is larger than the minimal NBD and requires a more extended conformation for high-affinity interaction.
The formation of a defined 2:2 complex suggests a specific structural arrangement critical for IKK function.
These findings provide a deeper understanding of the molecular basis for IKK-NEMO complex assembly and NF-kappaB regulation.