2V4H

NEMO CC2-LZ domain - 1D5 DARPin complex

2V4H の概要

• エントリーDOI: 10.2210/pdb2v4h/pdb
• 分子名称: NF-KAPPA-B ESSENTIAL MODULATOR, 1D5 DARPIN (3 entities in total)
• 機能のキーワード: transcription, metal-binding, nemo - ikk gamma - nfkb pathway -darpin, transcription regulation
• 由来する生物種: MUS MUSCULUS (HOUSE MOUSE); 詳細
• 細胞内の位置: Cytoplasm : O88522
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 55640.25

構造登録者

Grubisha, O.,Duquerroy, S.,Cordier, F.,Haouz, A.,Delepierre, M.,Veron, M.,Agou, F. (登録日: 2008-09-22, 公開日: 2009-11-03, 最終更新日: 2023-12-13)

主引用文献

Grubisha, O.,Kaminska, M.,Duquerroy, S.,Fontan, E.,Cordier, F.,Haouz, A.,Raynal, B.,Chiaravalli, J.,Delepierre, M.,Israel, A.,Veron, M.,Agou, F.
Darpin-Assisted Crystallography of the Cc2-Lz Domain of Nemo Reveals a Coupling between Dimerization and Ubiquitin-Binding.
J.Mol.Biol., 395:89-, 2010

PubMed Abstract: NEMO is an integral part of the IkappaB kinase complex and serves as a molecular switch by which the NF-kappaB signaling pathway can be regulated. Oligomerization and polyubiquitin (poly-Ub) binding, mediated through the regulatory CC2-LZ domain, were shown to be key features governing NEMO function, but the relationship between these two activities remains unclear. In this study, we solved the structure of this domain in complex with a designed ankyrin repeat protein, which helps its crystallization. We generated several NEMO mutants in this domain, including those associated with human diseases incontinentia pigmenti and immunodeficiency with or without anhidrotic ectodermal dysplasia. Analytical ultracentrifugation and thermal denaturation experiments were used to evaluate the dimerization properties of these mutants. A fluorescence-based assay was developed, as well, to quantify the interaction to monoubiquitin and poly-Ub chains. Moreover, the effect of these mutations was investigated for the full-length protein. We show that a proper folding of the ubiquitin-binding domain, termed NOA/UBAN/NUB, into a stable coiled-coil dimer is required but not sufficient for efficient interaction with poly-Ub. In addition, we show that binding to poly-Ub and, to a lesser extent, to monoubiquitin increases the stability of the NOA coiled-coil dimer. Collectively, these data provide structural insights into how several pathological mutations within and outside of the CC2-LZ's NOA ubiquitin binding site affect IkappaB kinase activation in the NF-kappaB signaling pathway.

PubMed: 19854204
DOI: 10.1016/J.JMB.2009.10.018

実験手法

X-RAY DIFFRACTION (2.9 Å)