6ESP

Proteome-wide analysis of phospho-regulated PDZ domain interactions

6ESP の概要

• エントリーDOI: 10.2210/pdb6esp/pdb
• NMR情報: BMRB: 34190
• 分子名称: Protein scribble homolog (1 entity in total)
• 機能のキーワード: pdz, phosphorylation, scribble, protein binding
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 12217.79

構造登録者

Chi, N.C. (登録日: 2017-10-24, 公開日: 2018-09-05, 最終更新日: 2024-05-15)

主引用文献

Sundell, G.N.,Arnold, R.,Ali, M.,Naksukpaiboon, P.,Orts, J.,Guntert, P.,Chi, C.N.,Ivarsson, Y.
Proteome-wide analysis of phospho-regulated PDZ domain interactions.
Mol. Syst. Biol., 14:e8129-e8129, 2018

PubMed Abstract: A key function of reversible protein phosphorylation is to regulate protein-protein interactions, many of which involve short linear motifs (3-12 amino acids). Motif-based interactions are difficult to capture because of their often low-to-moderate affinities. Here, we describe phosphomimetic proteomic peptide-phage display, a powerful method for simultaneously finding motif-based interaction and pinpointing phosphorylation switches. We computationally designed an oligonucleotide library encoding human C-terminal peptides containing known or predicted Ser/Thr phosphosites and phosphomimetic variants thereof. We incorporated these oligonucleotides into a phage library and screened the PDZ (PSD-95/Dlg/ZO-1) domains of Scribble and DLG1 for interactions potentially enabled or disabled by ligand phosphorylation. We identified known and novel binders and characterized selected interactions through microscale thermophoresis, isothermal titration calorimetry, and NMR We uncover site-specific phospho-regulation of PDZ domain interactions, provide a structural framework for how PDZ domains accomplish phosphopeptide binding, and discuss ligand phosphorylation as a switching mechanism of PDZ domain interactions. The approach is readily scalable and can be used to explore the potential phospho-regulation of motif-based interactions on a large scale.

PubMed: 30126976
DOI: 10.15252/msb.20178129

実験手法

SOLUTION NMR