7NAO

Human PA28-20S proteasome complex

7NAO の概要

• エントリーDOI: 10.2210/pdb7nao/pdb
• EMDBエントリー: 24275 24276 24277 24278
• 分子名称: Proteasome subunit alpha type-2, Proteasome subunit beta type-2, Proteasome subunit beta type-5, ... (16 entities in total)
• 機能のキーワード: proteasome, 20s, pa28, hydrolase
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 35
• 化学式量合計: 957654.25

構造登録者

Zhao, J.,Makhija, S.,Huang, B.,Cheng, Y. (登録日: 2021-06-22, 公開日: 2022-11-02, 最終更新日: 2024-06-05)

主引用文献

Zhao, J.,Makhija, S.,Zhou, C.,Zhang, H.,Wang, Y.,Muralidharan, M.,Huang, B.,Cheng, Y.
Structural insights into the human PA28-20S proteasome enabled by efficient tagging and purification of endogenous proteins.
Proc.Natl.Acad.Sci.USA, 119:e2207200119-e2207200119, 2022

PubMed Abstract: The ability to produce folded and functional proteins is a necessity for structural biology and many other biological sciences. This task is particularly challenging for numerous biomedically important targets in human cells, including membrane proteins and large macromolecular assemblies, hampering mechanistic studies and drug development efforts. Here we describe a method combining CRISPR-Cas gene editing and fluorescence-activated cell sorting to rapidly tag and purify endogenous proteins in HEK cells for structural characterization. We applied this approach to study the human proteasome from HEK cells and rapidly determined cryogenic electron microscopy structures of major proteasomal complexes, including a high-resolution structure of intact human PA28αβ-20S. Our structures reveal that PA28 with a subunit stoichiometry of 3α/4β engages tightly with the 20S proteasome. Addition of a hydrophilic peptide shows that polypeptides entering through PA28 are held in the antechamber of 20S prior to degradation in the proteolytic chamber. This study provides critical insights into an important proteasome complex and demonstrates key methodologies for the tagging of proteins from endogenous sources.

PubMed: 35858375
DOI: 10.1073/pnas.2207200119

実験手法

ELECTRON MICROSCOPY (2.9 Å)