8EFW

Structure of SdeA DUB Domain disulfide crosslinked with Ubiquitin

8EFW の概要

• エントリーDOI: 10.2210/pdb8efw/pdb
• 分子名称: SdeA, Ubiquitin (2 entities in total)
• 機能のキーワード: complex, deubiquitinase, hydrolase, hydrolase-signaling protein complex, hydrolase/signaling protein
• 由来する生物種: Legionella pneumophila; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 30325.39

構造登録者

Negron Teron, K.N.,Das, C. (登録日: 2022-09-09, 公開日: 2023-09-20, 最終更新日: 2024-10-30)

主引用文献

Negron Teron, K.I.,Das, C.
Cocrystallization of ubiquitin-deubiquitinase complexes through disulfide linkage.
Acta Crystallogr D Struct Biol, 79:1044-1055, 2023

PubMed Abstract: Structural characterization of the recognition of ubiquitin (Ub) by deubiquitinases (DUBs) has largely relied on covalent complexation of the DUB through its catalytic cysteine with a Ub C-terminal electrophile. The Ub electrophiles are accessed through intein chemistry in conjunction with chemical synthesis. Here, it was asked whether DUB-Ub covalent complexes could instead be accessed by simpler disulfide chemistry using a Ub cysteine mutant in which the last glycine has been replaced with a cysteine. The Ub cysteine mutant displayed a wide variability in disulfide formation across a panel of eukaryotic and prokaryotic DUBs, with some showing no detectable reaction while others robustly produced a disulfide complex. Using this approach, two disulfide-linked ubiquitin-bound complexes were crystallized, one involving the Legionella pneumophila effector SdeA DUB and the other involving the Orientia effector OtDUB. These DUBs had previously been crystallized in Ub-bound forms using the C-terminal electrophile strategy and noncovalent complexation, respectively. While the disulfide-linked SdeA DUB-Ub complex crystallized as expected, in the OtDUB complex the disulfide bond to the Ub mutant involved a cysteine that differed from the catalytic cysteine. Disulfide formation with the SdeA DUB catalytic cysteine was accompanied by local distortion of the helix carrying the active-site cysteine, whereas OtDUB reacted with the Ub mutant using a surface-exposed cysteine.

PubMed: 37877948
DOI: 10.1107/S2059798323008501

実験手法

X-RAY DIFFRACTION (2.81 Å)