6FHP

DAIP in complex with a C-terminal fragment of thermolysin

6FHP の概要

• エントリーDOI: 10.2210/pdb6fhp/pdb
• 関連するPDBエントリー: 5fzp
• 分子名称: Dispase autolysis-inducing protein, Thermolysin (3 entities in total)
• 機能のキーワード: dispase autolysis-inducing protein; neutral metalloproteases; protease twisting; conformation analysis, antimicrobial protein
• 由来する生物種: Streptomyces mobaraensis (Streptoverticillium mobaraense); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 84336.84

構造登録者

Schmelz, S.,Fiebig, D.,Fuchsbauer, H.L.,Blankenfeldt, W.,Scrima, A. (登録日: 2018-01-15, 公開日: 2018-09-12, 最終更新日: 2024-11-20)

主引用文献

Fiebig, D.,Storka, J.,Roeder, M.,Meyners, C.,Schmelz, S.,Blankenfeldt, W.,Scrima, A.,Kolmar, H.,Fuchsbauer, H.L.
Destructive twisting of neutral metalloproteases: the catalysis mechanism of the Dispase autolysis-inducing protein from Streptomyces mobaraensis DSM 40487.
FEBS J., 285:4246-4264, 2018

PubMed Abstract: The Dispase autolysis-inducing protein (DAIP) is produced by Streptomyces mobaraensis to disarm neutral metalloproteases by decomposition. The absence of a catalytic protease domain led to the assumption that the seven-bladed β-propeller protein DAIP causes structural modifications, thereby triggering autolysis. Determination of protein complexes consisting of DAIP and thermolysin or DAIP and a nonfunctional E138A bacillolysin variant supported this postulation. Protein twisting was indicated by DAIP-mediated inhibition of thermolysin while bacillolysin underwent immediate autolysis under the same conditions. Interestingly, an increase in SYPRO orange fluorescence allowed tracking of the fast degradation process. Similarly rapid autolysis of thermolysin mediated by DAIP was only observed upon the addition of amphiphilic compounds, which probably amplify the induced structural changes. DAIP further caused degradation of FITC-labeled E138A bacillolysin by trypsin, as monitored by a linear decrease in fluorescence polarization. The kinetic model, calculated from the obtained data, suggested a three-step mechanism defined by (a) fast DAIP-metalloprotease complex formation, (b) slower DAIP-mediated protein twisting, and (c) fragmentation. These results were substantiated by crystallized DAIP attached to a C-terminal helix fragment of thermolysin. Structural superposition of the complex with thermolysin is indicative of a conformational change upon binding to DAIP. Importantly, the majority of metalloproteases, also including homologs from various pathogens, are highly conserved at the autolysis-prone peptide bonds, suggesting their susceptibility to DAIP-mediated decomposition, which may offer opportunities for pharmaceutical applications. DATABASES: The atomic coordinates and structure factors (PDB ID: 6FHP) have been deposited in the Protein Data Bank (http://www.pdb.org/). ENZYMES: Aureolysin, EC 3.4.24.29; bacillolysin (Dispase, Gentlyase), EC 3.4.24.28; lasB (elastase), EC 3.4.24.4; subtilisin, EC 3.4.21.62; thermolysin, EC 3.4.24.27; transglutaminase, EC 2.3.2.13; trypsin, EC 3.4.21.4; vibriolysin (hemagglutinin(HA)/protease), EC 3.4.24.25.

PubMed: 30171661
DOI: 10.1111/febs.14647

実験手法

X-RAY DIFFRACTION (1.703 Å)