5YWR

Crystal Structure of RING E3 ligase ZNRF1 in complex with Ube2N (Ubc13)

5YWR の概要

• エントリーDOI: 10.2210/pdb5ywr/pdb
• 分子名称: Ubiquitin-conjugating enzyme E2 N, E3 ubiquitin-protein ligase ZNRF1, TRIETHYLENE GLYCOL, ... (6 entities in total)
• 機能のキーワード: ubiquitin e3 ligase, znrf1, ubiquitin conjugating e2, ube2n, ubc13, signaling 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: 27574.73

構造登録者

Behera, A.P.,Naskar, P.,Datta, A.B. (登録日: 2017-11-30, 公開日: 2018-06-06, 最終更新日: 2024-03-27)

主引用文献

Behera, A.P.,Naskar, P.,Agarwal, S.,Banka, P.A.,Poddar, A.,Datta, A.B.
Structural insights into the nanomolar affinity of RING E3 ligase ZNRF1 for Ube2N and its functional implications.
Biochem. J., 475:1569-1582, 2018

PubMed Abstract: RING (eally nteresting ew ene) domains in ubiquitin RING E3 ligases exclusively engage ubiquitin (Ub)-loaded E2s to facilitate ubiquitination of their substrates. Despite such specificity, all RINGs characterized till date bind unloaded E2s with dissociation constants (s) in the micromolar to the sub-millimolar range. Here, we show that the RING domain of E3 ligase ZNRF1, an essential E3 ligase implicated in diverse cellular pathways, binds Ube2N with a of ∼50 nM. This high-affinity interaction is exclusive for Ube2N as ZNRF1 interacts with Ube2D2 with a of ∼1 µM, alike few other E3s. The crystal structure of ZNRF1 C-terminal domain in complex with Ube2N coupled with mutational analyses reveals the molecular basis of this unusual affinity. We further demonstrate that the ubiquitination efficiency of ZNRF1 : E2 pairs correlates with their affinity. Intriguingly, as a consequence of its high E2 affinity, an excess of ZNRF1 inhibits Ube2N-mediated ubiquitination at concentrations ≥500 nM instead of showing enhanced ubiquitination. This suggests a novel mode of activity regulation of E3 ligases and emphasizes the importance of E3-E2 balance for the optimum activity. Based on our results, we propose that overexpression-based functional analyses on E3 ligases such as ZNRF1 must be approached with caution as enhanced cellular levels might result in aberrant modification activity.

PubMed: 29626159
DOI: 10.1042/BCJ20170909

実験手法

X-RAY DIFFRACTION (1.47 Å)