5OE7

Structure of OTULIN bound to the Met1-linked diubiquitin activity probe

5OE7 の概要

• エントリーDOI: 10.2210/pdb5oe7/pdb
• 分子名称: Ubiquitin thioesterase otulin, Polyubiquitin-C (2 entities in total)
• 機能のキーワード: deubiquitinase complex otu, hydrolase
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Cytoplasm : Q96BN8; Ubiquitin: Cytoplasm : P0CG48
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 48219.28

構造登録者

Elliott, P.R.,Komander, D. (登録日: 2017-07-07, 公開日: 2017-09-27, 最終更新日: 2024-01-17)

主引用文献

Weber, A.,Elliott, P.R.,Pinto-Fernandez, A.,Bonham, S.,Kessler, B.M.,Komander, D.,El Oualid, F.,Krappmann, D.
A Linear Diubiquitin-Based Probe for Efficient and Selective Detection of the Deubiquitinating Enzyme OTULIN.
Cell Chem Biol, 24:1299-1313.e7, 2017

PubMed Abstract: The methionine 1 (M1)-specific deubiquitinase (DUB) OTULIN acts as a negative regulator of nuclear factor κB signaling and immune homeostasis. By replacing Gly76 in distal ubiquitin (Ub) by dehydroalanine we designed the diubiquitin (diUb) activity-based probe Ub-Ub (OTULIN activity-based probe [ABP]) that couples to the catalytic site of OTULIN and thereby captures OTULIN in its active conformation. The OTULIN ABP displays high selectivity for OTULIN and does not label other M1-cleaving DUBs, including CYLD. The only detectable cross-reactivities were the labeling of USP5 (Isopeptidase T) and an ATP-dependent assembly of polyOTULIN ABP chains via Ub-activating E1 enzymes. Both cross-reactivities were abolished by the removal of the C-terminal Gly in the ABP's proximal Ub, yielding the specific OTULIN probe Ub-Ub (OTULIN ABPΔG76). Pull-downs demonstrate that substrate-bound OTULIN associates with the linear ubiquitin chain assembly complex (LUBAC). Thus, we present a highly selective ABP for OTULIN that will facilitate studying the cellular function of this essential DUB.

PubMed: 28919039
DOI: 10.1016/j.chembiol.2017.08.006

実験手法

X-RAY DIFFRACTION (2.95 Å)