6YEK

Crystal structure of human NEMO apo form

6YEK の概要

• エントリーDOI: 10.2210/pdb6yek/pdb
• 関連するPDBエントリー: 6XX0
• 分子名称: Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase gamma, isoform CRA_b (2 entities in total)
• 機能のキーワード: cell signaling, nf-kb pathway, protein binding
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 20663.59

構造登録者

Garcia-Pardo, J.,Akutsu, M.,Busse, P.,Skenderovic, A.,Maculins, T.,Dikic, I. (登録日: 2020-03-25, 公開日: 2021-03-03, 最終更新日: 2024-01-24)

主引用文献

Maculins, T.,Garcia-Pardo, J.,Skenderovic, A.,Gebel, J.,Putyrski, M.,Vorobyov, A.,Busse, P.,Varga, G.,Kuzikov, M.,Zaliani, A.,Rahighi, S.,Schaeffer, V.,Parnham, M.J.,Sidhu, S.S.,Ernst, A.,Dotsch, V.,Akutsu, M.,Dikic, I.
Discovery of Protein-Protein Interaction Inhibitors by Integrating Protein Engineering and Chemical Screening Platforms.
Cell Chem Biol, 27:1441-1451.e7, 2020

PubMed Abstract: Protein-protein interactions (PPIs) govern intracellular life, and identification of PPI inhibitors is challenging. Roadblocks in assay development stemming from weak binding affinities of natural PPIs impede progress in this field. We postulated that enhancing binding affinity of natural PPIs via protein engineering will aid assay development and hit discovery. This proof-of-principle study targets PPI between linear ubiquitin chains and NEMO UBAN domain, which activates NF-κB signaling. Using phage display, we generated ubiquitin variants that bind to the functional UBAN epitope with high affinity, act as competitive inhibitors, and structurally maintain the existing PPI interface. When utilized in assay development, variants enable generation of robust cell-based assays for chemical screening. Top compounds identified using this approach directly bind to UBAN and dampen NF-κB signaling. This study illustrates advantages of integrating protein engineering and chemical screening in hit identification, a development that we anticipate will have wide application in drug discovery.

PubMed: 32726587
DOI: 10.1016/j.chembiol.2020.07.010

実験手法

X-RAY DIFFRACTION (3.2 Å)