5KYC

Crystal structure of USP7 catalytic domain [V302K] mutant in complex with ubiquitin (malonate bound)

5KYC の概要

• エントリーDOI: 10.2210/pdb5kyc/pdb
• 関連するPDBエントリー: 5KYB 5KYD 5KYE 5KYF
• 分子名称: Ubiquitin carboxyl-terminal hydrolase 7, Polyubiquitin-B, MALONIC ACID, ... (5 entities in total)
• 機能のキーワード: usp7 catalytic domain, deubiquitinase, v302k mutation, hydrolase
• 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: 49483.01

構造登録者

Rouge, L.,Ozen, A. (登録日: 2016-07-21, 公開日: 2017-08-09, 最終更新日: 2024-10-16)

主引用文献

Ozen, A.,Rouge, L.,Bashore, C.,Hearn, B.R.,Skelton, N.J.,Dueber, E.C.
Selectively Modulating Conformational States of USP7 Catalytic Domain for Activation.
Structure, 26:72-84.e7, 2018

PubMed Abstract: Ubiquitin-specific protease 7 (USP7) deubiquitinase activity is controlled by a number of regulatory factors, including stimulation by intramolecular accessory domains. Alone, the USP7 catalytic domain (USP7cd) shows limited activity and apo USP7cd crystal structures reveal a disrupted catalytic triad. By contrast, ubiquitin-conjugated USP7cd structures demonstrate the canonical cysteine protease active-site geometry; however, the structural features of the USP7cd that stabilize the inactive conformation and the mechanism of transition between inactive and active states remain unclear. Here we use comparative structural analyses, molecular dynamics simulations, and in silico sequence re-engineering via directed sampling by RosettaDesign to identify key molecular determinants of USP7cd activation and successfully engineer USP7cd for improved activity. Full kinetic analysis and multiple X-ray crystal structures of our designs indicate that electrostatic interactions in the distal "switching loop" region and local packing in the hydrophobic core mediate subtle but significant conformational changes that modulate USP7cd activation.

PubMed: 29249604
DOI: 10.1016/j.str.2017.11.010

実験手法

X-RAY DIFFRACTION (1.426 Å)