4OYL

Humicola insolens cutinase in complex with mono-ethylphosphate

4OYL の概要

• エントリーDOI: 10.2210/pdb4oyl/pdb
• 関連するPDBエントリー: 4OYY
• 分子名称: Cutinase (2 entities in total)
• 機能のキーワード: hydrolase
• 由来する生物種: Humicola insolens
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 61217.90

構造登録者

Dauter, Z.D.,Brzozowski, A.M.,Turkenburg, J.P.,Wilson, K.S. (登録日: 2014-02-12, 公開日: 2014-06-25, 最終更新日: 2023-12-27)

主引用文献

Kold, D.,Dauter, Z.,Laustsen, A.K.,Brzozowski, A.M.,Turkenburg, J.P.,Nielsen, A.D.,Kolds, H.,Petersen, E.,Schitt, B.,De Maria, L.,Wilson, K.S.,Svendsen, A.,Wimmer, R.
Thermodynamic and structural investigation of the specific SDS binding of Humicola insolens cutinase.
Protein Sci., 23:1023-1035, 2014

PubMed Abstract: The interaction of lipolytic enzymes with anionic surfactants is of great interest with respect to industrially produced detergents. Here, we report the interaction of cutinase from the thermophilic fungus Humicola insolens with the anionic surfactant SDS, and show the enzyme specifically binds a single SDS molecule under nondenaturing concentrations. Protein interaction with SDS was investigated by NMR, ITC and molecular dynamics simulations. The NMR resonances of the protein were assigned, with large stretches of the protein molecule not showing any detectable resonances. SDS is shown to specifically interact with the loops surrounding the catalytic triad with medium affinity (Ka ≈ 10(5) M(-1) ). The mode of binding is closely similar to that seen previously for binding of amphiphilic molecules and substrate analogues to cutinases, and hence SDS acts as a substrate mimic. In addition, the structure of the enzyme has been solved by X-ray crystallography in its apo form and after cocrystallization with diethyl p-nitrophenyl phosphate (DNPP) leading to a complex with monoethylphosphate (MEP) esterified to the catalytically active serine. The enzyme has the same fold as reported for other cutinases but, unexpectedly, esterification of the active site serine is accompanied by the ethylation of the active site histidine which flips out from its usual position in the triad.

PubMed: 24832484
DOI: 10.1002/pro.2489

実験手法

X-RAY DIFFRACTION (2.05 Å)