5D0T

Yeast 20S proteasome beta5-D166N mutant in complex with MG132

5D0T の概要

• エントリーDOI: 10.2210/pdb5d0t/pdb
• 関連するPDBエントリー: 5CZ4
• 関連するBIRD辞書のPRD_ID: PRD_001210
• 分子名称: Proteasome subunit alpha type-2, Proteasome subunit beta type-4, Proteasome subunit beta type-5, ... (18 entities in total)
• 機能のキーワード: hydrolase-hydrolase inhibitor complex, proteasome, mutant, inhibitor, binding analysis, hydrolase/hydrolase inhibitor
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c); 詳細
• タンパク質・核酸の鎖数: 28
• 化学式量合計: 733368.51

構造登録者

Huber, E.M.,Groll, M. (登録日: 2015-08-03, 公開日: 2016-03-23, 最終更新日: 2024-01-10)

主引用文献

Huber, E.M.,Heinemeyer, W.,Li, X.,Arendt, C.S.,Hochstrasser, M.,Groll, M.
A unified mechanism for proteolysis and autocatalytic activation in the 20S proteasome.
Nat Commun, 7:10900-10900, 2016

PubMed Abstract: Biogenesis of the 20S proteasome is tightly regulated. The N-terminal propeptides protecting the active-site threonines are autocatalytically released only on completion of assembly. However, the trigger for the self-activation and the reason for the strict conservation of threonine as the active site nucleophile remain enigmatic. Here we use mutagenesis, X-ray crystallography and biochemical assays to suggest that Lys33 initiates nucleophilic attack of the propeptide by deprotonating the Thr1 hydroxyl group and that both residues together with Asp17 are part of a catalytic triad. Substitution of Thr1 by Cys disrupts the interaction with Lys33 and inactivates the proteasome. Although a Thr1Ser mutant is active, it is less efficient compared with wild type because of the unfavourable orientation of Ser1 towards incoming substrates. This work provides insights into the basic mechanism of proteolysis and propeptide autolysis, as well as the evolutionary pressures that drove the proteasome to become a threonine protease.

PubMed: 26964885
DOI: 10.1038/ncomms10900

実験手法

X-RAY DIFFRACTION (2.6 Å)