6XXC

Ternary complex of 14-3-3 sigma (C38N), Estrogen Related Receptor gamma (DBD) phosphopeptide, and disulfide PPI stabilizer 4

6XXC の概要

• エントリーDOI: 10.2210/pdb6xxc/pdb
• 分子名称: 14-3-3 protein sigma, Estrogen Related Receptor gamma phosphopeptide, MAGNESIUM ION, ... (6 entities in total)
• 機能のキーワード: 14-3-3, erry phosphopeptide, disulfide, peptide binding protein
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 28074.89

構造登録者

Somsen, B.A.,Ottmann, C. (登録日: 2020-01-27, 公開日: 2020-11-18, 最終更新日: 2024-01-24)

主引用文献

Sijbesma, E.,Somsen, B.A.,Miley, G.P.,Leijten-van de Gevel, I.A.,Brunsveld, L.,Arkin, M.R.,Ottmann, C.
Fluorescence Anisotropy-Based Tethering for Discovery of Protein-Protein Interaction Stabilizers.
Acs Chem.Biol., 15:3143-3148, 2020

PubMed Abstract: Protein-protein interaction (PPI) networks are fundamental for cellular processes. Small-molecule PPI enhancers have been shown to be powerful tools to fundamentally study PPIs and as starting points for potential new therapeutics. Yet, systematic approaches for their discovery are not widely available, and the design prerequisites of "molecular glues" are poorly understood. Covalent fragment-based screening can identify chemical starting points for these enhancers at specific sites in PPI interfaces. We recently reported a mass spectrometry-based disulfide-trapping (tethering) approach for a cysteine residue in the hub protein 14-3-3, an important regulator of phosphorylated client proteins. Here, we invert the strategy and report the development of a functional read-out for systematic identification of PPI enhancers based on fluorescence anisotropy (FA-tethering) with the reactive handle now on a client-derived peptide. Using the DNA-binding domain of the nuclear receptor Estrogen Related Receptor gamma (ERRγ), we target a native cysteine positioned at the 14-3-3 PPI interface and identify several fragments that form a disulfide bond to ERRγ and stabilize the complex up to 5-fold. Crystallography indicates that fragments bind in a pocket comprised of 14-3-3 and the ERRγ phosphopeptide. FA-tethering presents a streamlined methodology to discover molecular glues for protein complexes.

PubMed: 33196173
DOI: 10.1021/acschembio.0c00646

実験手法

X-RAY DIFFRACTION (1.3 Å)