6UAI

Imidazole-triggered RAS-specific subtilisin SUBT_BACAM complexed with YSAM peptide

6UAI の概要

• エントリーDOI: 10.2210/pdb6uai/pdb
• 関連するPDBエントリー: 6U9L
• 分子名称: SUBTILISIN BPN', YSAM peptide, SODIUM ION, ... (7 entities in total)
• 機能のキーワード: engineered protease, hydrolase
• 由来する生物種: Bacillus amyloliquefaciens; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 27362.88

構造登録者

Toth, E.A.,Bryan, P.N.,Orban, J. (登録日: 2019-09-10, 公開日: 2020-09-16, 最終更新日: 2024-11-20)

主引用文献

Chen, Y.,Toth, E.A.,Ruan, B.,Choi, E.J.,Simmerman, R.,Chen, Y.,He, Y.,Wang, R.,Godoy-Ruiz, R.,King, H.,Custer, G.,Travis Gallagher, D.,Rozak, D.A.,Solomon, M.,Muro, S.,Weber, D.J.,Orban, J.,Fuerst, T.R.,Bryan, P.N.
Engineering subtilisin proteases that specifically degrade active RAS.
Commun Biol, 4:299-299, 2021

PubMed Abstract: We describe the design, kinetic properties, and structures of engineered subtilisin proteases that degrade the active form of RAS by cleaving a conserved sequence in switch 2. RAS is a signaling protein that, when mutated, drives a third of human cancers. To generate high specificity for the RAS target sequence, the active site was modified to be dependent on a cofactor (imidazole or nitrite) and protease sub-sites were engineered to create a linkage between substrate and cofactor binding. Selective proteolysis of active RAS arises from a 2-step process wherein sub-site interactions promote productive binding of the cofactor, enabling cleavage. Proteases engineered in this way specifically cleave active RAS in vitro, deplete the level of RAS in a bacterial reporter system, and also degrade RAS in human cell culture. Although these proteases target active RAS, the underlying design principles are fundamental and will be adaptable to many target proteins.

PubMed: 33674772
DOI: 10.1038/s42003-021-01818-7

実験手法

X-RAY DIFFRACTION (1.2 Å)