9SDX

Structure of RBR binding E2 variant crosslinked with NEDD8-CUL5-RBX2 bound ARIH2 and Ub

9SDX の概要

• エントリーDOI: 10.2210/pdb9sdx/pdb
• EMDBエントリー: 54793
• 分子名称: NEDD8, RING-box protein 2, Ubiquitin, ... (8 entities in total)
• 機能のキーワード: complex, ligase
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 197840.53

構造登録者

Schulman, B.A.,Du, J. (登録日: 2025-08-15, 公開日: 2025-12-24, 最終更新日: 2026-01-14)

主引用文献

Du, J.,Andree, G.A.,Horn-Ghetko, D.,Stier, L.,Singh, J.,Kostrhon, S.,Kiss, L.,Mann, M.,Sidhu, S.S.,Schulman, B.A.
E2 variants for probing E3 ubiquitin ligase activities.
Proc.Natl.Acad.Sci.USA, 123:e2524899122-e2524899122, 2026

PubMed Abstract: E3 ligases partner with E2 enzymes to regulate vast eukaryotic biology. The hierarchical nature of these pairings, with >600 E3s and ~40 E2s in humans, necessitates that E2s cofunction with numerous different E3s. Here, focusing on E3s in the RING-between-RING (RBR) family and their partner UBE2L3 and UBE2D-family E2s, we report an approach to interrogate selected pathways. We screened phage-displayed libraries of structure-based E2 variants (E2Vs) to discover enzymes with enhanced affinity and specificity toward half of all RBR E3 ligases (ARIH1, ARIH2, ANKIB1, CUL9, HOIL1, HOIP, and RNF14). Collectively, these E2Vs allowed distinguishing actions of different cofunctioning E3s, obtaining high-resolution cryogenic Electron Microscopy (cryo-EM) structures of an RBR E3 in the context of a substrate-bound multiprotein complex, and profiling an endogenous RBR E3 response to an extracellular stimulus. Overall, we anticipate that E2V technology will be a generalizable tool to enable in-depth mechanistic and structural analysis of E3 ligase functions, and mapping their activity states and protein partners in cellular signaling cascades.

PubMed: 41481455
DOI: 10.1073/pnas.2524899122

実験手法

ELECTRON MICROSCOPY (2.97 Å)