1I19

CRYSTAL STRUCTURE OF CHOLESTEROL OXIDASE FROM B.STEROLICUM

1I19 の概要

• エントリーDOI: 10.2210/pdb1i19/pdb
• 分子名称: CHOLESTEROL OXIDASE, 1,2-ETHANEDIOL, FLAVIN-ADENINE DINUCLEOTIDE, ... (6 entities in total)
• 機能のキーワード: mix alpha beta, covalent fad, flavoenzyme, oxidoreductase
• 由来する生物種: Brevibacterium sterolicum
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 125767.88

構造登録者

Coulombe, R.,Yue, K.Q.,Ghisla, S.,Vrielink, A. (登録日: 2001-01-31, 公開日: 2001-08-08, 最終更新日: 2024-11-13)

主引用文献

Coulombe, R.,Yue, K.Q.,Ghisla, S.,Vrielink, A.
Oxygen access to the active site of cholesterol oxidase through a narrow channel is gated by an Arg-Glu pair.
J.Biol.Chem., 276:30435-30441, 2001

PubMed Abstract: Cholesterol oxidase is a monomeric flavoenzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one. Two forms of the enzyme are known, one containing the cofactor non-covalently bound to the protein and one in which the cofactor is covalently linked to a histidine residue. The x-ray structure of the enzyme from Brevibacterium sterolicum containing covalently bound FAD has been determined and refined to 1.7-A resolution. The active site consists of a cavity sealed off from the exterior of the protein. A model for the steroid substrate, cholesterol, can be positioned in the pocket revealing the structural factors that result in different substrate binding affinities between the two known forms of the enzyme. The structure suggests that Glu(475), located at the active site cavity, may act as the base for both the oxidation and the isomerization steps of the catalytic reaction. A water-filled channel extending toward the flavin moiety, inside the substrate-binding cavity, may act as the entry point for molecular oxygen for the oxidative half-reaction. An arginine and a glutamate residue at the active site, found in two conformations are proposed to control oxygen access to the cavity from the channel. These concerted side chain movements provide an explanation for the biphasic mode of reaction with dioxygen and the ping-pong kinetic mechanism exhibited by the enzyme.

PubMed: 11397813
DOI: 10.1074/jbc.M104103200

実験手法

X-RAY DIFFRACTION (1.7 Å)