High-affinity interaction between IKKβ and NEMO

Yu-Chih Lo, Upendra Maddineni, Jee Y. Chung, Rebecca L. Rich, David G. Myszka, Hao Wu

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The Ser/Thr-specific IκB kinase (IKK), which comprises IKKα or IKKβ and the regulatory protein NEMO, is at the bottleneck for NF-κB activation. IKK activity relies on interaction between NEMO and IKKα or IKKβ. A conserved region in the C-terminal tail of IKKβ or IKKα (NEMO-binding domain, NBD, residues 734-745 of IKKβ) is important for interaction with NEMO. Here we show that the NBD peptide of IKKβ is not sufficient for interaction with NEMO. Instead, a longer region of the IKKβ C-terminal region provides high affinity for NEMO. Quantitative measurements using surface plasmon resonance and isothermal titration calorimetry confirm the differential affinities of these interactions and provide insight into the kinetic and thermodynamic behaviors of the interactions. Biochemical characterization using multiangle light scattering (MALS) coupled with refractive index shows that the longer IKKβ C-terminal region forms a 2:2 stoichiometirc complex with NEMO.

Original languageEnglish
Pages (from-to)3109-3116
Number of pages8
JournalBiochemistry
Volume47
Issue number10
DOIs
Publication statusPublished - 2008 Mar 11

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Refractometry
Calorimetry
Surface Plasmon Resonance
Surface plasmon resonance
Titration
Thermodynamics
Light scattering
Refractive index
Phosphotransferases
Chemical activation
Light
Kinetics
Proteins
IKK NBD peptide

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Lo, Y-C., Maddineni, U., Chung, J. Y., Rich, R. L., Myszka, D. G., & Wu, H. (2008). High-affinity interaction between IKKβ and NEMO. Biochemistry, 47(10), 3109-3116. https://doi.org/10.1021/bi702312c
Lo, Yu-Chih ; Maddineni, Upendra ; Chung, Jee Y. ; Rich, Rebecca L. ; Myszka, David G. ; Wu, Hao. / High-affinity interaction between IKKβ and NEMO. In: Biochemistry. 2008 ; Vol. 47, No. 10. pp. 3109-3116.
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Lo, Y-C, Maddineni, U, Chung, JY, Rich, RL, Myszka, DG & Wu, H 2008, 'High-affinity interaction between IKKβ and NEMO', Biochemistry, vol. 47, no. 10, pp. 3109-3116. https://doi.org/10.1021/bi702312c

High-affinity interaction between IKKβ and NEMO. / Lo, Yu-Chih; Maddineni, Upendra; Chung, Jee Y.; Rich, Rebecca L.; Myszka, David G.; Wu, Hao.

In: Biochemistry, Vol. 47, No. 10, 11.03.2008, p. 3109-3116.

Research output: Contribution to journalArticle

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Lo Y-C, Maddineni U, Chung JY, Rich RL, Myszka DG, Wu H. High-affinity interaction between IKKβ and NEMO. Biochemistry. 2008 Mar 11;47(10):3109-3116. https://doi.org/10.1021/bi702312c