4D13

Crystal structure of cofactor-free urate oxidase in complex with its 5-peroxoisourate intermediate (X-ray dose, 2.2 kGy)

4D13 の概要

• エントリーDOI: 10.2210/pdb4d13/pdb
• 関連するPDBエントリー: 4D12 4D17 4D19
• 分子名称: URICASE, 5-(HYDRO)PEROXOISOURATE, (4S)-2-METHYL-2,4-PENTANEDIOL, ... (4 entities in total)
• 機能のキーワード: oxidoreductase, cofactor-free oxidase
• 由来する生物種: ASPERGILLUS FLAVUS
• 細胞内の位置: Peroxisome: Q00511
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 34607.02

構造登録者

Bui, S.,Steiner, R.A. (登録日: 2014-05-01, 公開日: 2014-10-29, 最終更新日: 2024-05-08)

主引用文献

Bui, S.,von Stetten, D.,Jambrina, P.G.,Prange, T.,Colloc'h, N.,de Sanctis, D.,Royant, A.,Rosta, E.,Steiner, R.A.
Direct evidence for a peroxide intermediate and a reactive enzyme-substrate-dioxygen configuration in a cofactor-free oxidase.
Angew. Chem. Int. Ed. Engl., 53:13710-13714, 2014

PubMed Abstract: Cofactor-free oxidases and oxygenases promote and control the reactivity of O2 with limited chemical tools at their disposal. Their mechanism of action is not completely understood and structural information is not available for any of the reaction intermediates. Near-atomic resolution crystallography supported by in crystallo Raman spectroscopy and QM/MM calculations showed unambiguously that the archetypical cofactor-free uricase catalyzes uric acid degradation via a C5(S)-(hydro)peroxide intermediate. Low X-ray doses break specifically the intermediate C5-OO(H) bond at 100 K, thus releasing O2 in situ, which is trapped above the substrate radical. The dose-dependent rate of bond rupture followed by combined crystallographic and Raman analysis indicates that ionizing radiation kick-starts both peroxide decomposition and its regeneration. Peroxidation can be explained by a mechanism in which the substrate radical recombines with superoxide transiently produced in the active site.

PubMed: 25314114
DOI: 10.1002/anie.201405485

実験手法

X-RAY DIFFRACTION (1.3 Å)