7YPH

Open-spiral pentamer of the substrate-free Lon protease with a Y224S mutation

7YPH の概要

• エントリーDOI: 10.2210/pdb7yph/pdb
• EMDBエントリー: 34000
• 分子名称: Lon protease, PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER (2 entities in total)
• 機能のキーワード: lon protease, hydrolysis, aaa proteins, hydrolase
• 由来する生物種: Meiothermus taiwanensis
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 443438.95

構造登録者

Li, S.,Hsieh, K.Y.,Kuo, C.I.,Lee, S.H.,Ho, M.R.,Wang, C.H.,Zhang, K.,Chang, C.I. (登録日: 2022-08-03, 公開日: 2023-10-25, 最終更新日: 2023-11-29)

主引用文献

Li, S.,Hsieh, K.Y.,Kuo, C.I.,Lin, T.C.,Lee, S.H.,Chen, Y.R.,Wang, C.H.,Ho, M.R.,Ting, S.Y.,Zhang, K.,Chang, C.I.
A 5+1 assemble-to-activate mechanism of the Lon proteolytic machine.
Nat Commun, 14:7340-7340, 2023

PubMed Abstract: Many AAA+ (ATPases associated with diverse cellular activities) proteins function as protein or DNA remodelers by threading the substrate through the central pore of their hexameric assemblies. In this ATP-dependent translocating state, the substrate is gripped by the pore loops of the ATPase domains arranged in a universal right-handed spiral staircase organization. However, the process by which a AAA+ protein is activated to adopt this substrate-pore-loop arrangement remains unknown. We show here, using cryo-electron microscopy (cryo-EM), that the activation process of the Lon AAA+ protease may involve a pentameric assembly and a substrate-dependent incorporation of the sixth protomer to form the substrate-pore-loop contacts seen in the translocating state. Based on the structural results, we design truncated monomeric mutants that inhibit Lon activity by binding to the native pentamer and demonstrated that expressing these monomeric mutants in Escherichia coli cells containing functional Lon elicits specific phenotypes associated with lon deficiency, including the inhibition of persister cell formation. These findings uncover a substrate-dependent assembly process for the activation of a AAA+ protein and demonstrate a targeted approach to selectively inhibit its function within cells.

PubMed: 37957149
DOI: 10.1038/s41467-023-43035-2

実験手法

ELECTRON MICROSCOPY (3.68 Å)