5F5K

E.Coli GlpG Y205F mutant complexed with aldehyde inhibitor in DMPC/CHAPSO bicelle

5F5K の概要

• エントリーDOI: 10.2210/pdb5f5k/pdb
• 関連するPDBエントリー: 5F5B 5F5D 5F5G 5F5J
• 分子名称: Rhomboid protease GlpG, Peptidic derivative of Gurken: ACE-ARG-LYS-VAL-ARG-MET-ALA-aldehyde (3 entities in total)
• 機能のキーワード: glpg, rhomboid, intramembrane protease, bicelle, aldehyde inhibitor, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• 由来する生物種: Escherichia coli; 詳細
• 細胞内の位置: Cell inner membrane ; Multi-pass membrane protein . Cell membrane ; Multi-pass membrane protein : A0A0J2E248
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 24573.16

構造登録者

Urban, S.,Cho, S.,Dickey, S.W. (登録日: 2015-12-04, 公開日: 2016-02-10, 最終更新日: 2024-07-10)

主引用文献

Cho, S.,Dickey, S.W.,Urban, S.
Crystal Structures and Inhibition Kinetics Reveal a Two-Stage Catalytic Mechanism with Drug Design Implications for Rhomboid Proteolysis.
Mol.Cell, 61:329-340, 2016

PubMed Abstract: Intramembrane proteases signal by releasing proteins from the membrane, but despite their importance, their enzymatic mechanisms remain obscure. We probed rhomboid proteases with reversible, mechanism-based inhibitors that allow precise kinetic analysis and faithfully mimic the transition state structurally. Unexpectedly, inhibition by peptide aldehydes is non-competitive, revealing that in the Michaelis complex, substrate does not contact the catalytic center. Structural analysis in a membrane revealed that all extracellular loops of rhomboid make stabilizing interactions with substrate, but mainly through backbone interactions, explaining rhomboid's broad sequence selectivity. At the catalytic site, the tetrahedral intermediate lies covalently attached to the catalytic serine alone, with the oxyanion stabilized by unusual tripartite interactions with the side chains of H150, N154, and the backbone of S201. We also visualized unexpected substrate-enzyme interactions at the non-essential P2/P3 residues. These "extra" interactions foster potent rhomboid inhibition in living cells, thereby opening avenues for rational design of selective rhomboid inhibitors.

PubMed: 26805573
DOI: 10.1016/j.molcel.2015.12.022

実験手法

X-RAY DIFFRACTION (2.4 Å)