3VCP

The 2.2 Angstrom structure of Stc2 with proline bound in the active site

3VCP の概要

• エントリーDOI: 10.2210/pdb3vcp/pdb
• 関連するPDBエントリー: 3VCA
• 分子名称: Ring-hydroxylating dioxygenase, FE2/S2 (INORGANIC) CLUSTER, FE (III) ION, ... (5 entities in total)
• 機能のキーワード: rieske-type, mononuclear non-heme iron, n-demethylase, oxidoreductase
• 由来する生物種: Sinorhizobium meliloti (Ensifer meliloti)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 46772.25

構造登録者

Daughtry, K.D.,Xiao, Y.,Stoner-Ma, D.,Cho, E.,Orville, A.M.,Liu, P.,Allen, K.N. (登録日: 2012-01-04, 公開日: 2012-02-08, 最終更新日: 2023-09-13)

主引用文献

Daughtry, K.D.,Xiao, Y.,Stoner-Ma, D.,Cho, E.,Orville, A.M.,Liu, P.,Allen, K.N.
Quaternary Ammonium Oxidative Demethylation: X-ray Crystallographic, Resonance Raman, and UV-Visible Spectroscopic Analysis of a Rieske-Type Demethylase.
J.Am.Chem.Soc., 134:2823-2834, 2012

PubMed Abstract: Herein, the structure resulting from in situ turnover in a chemically challenging quaternary ammonium oxidative demethylation reaction was captured via crystallographic analysis and analyzed via single-crystal spectroscopy. Crystal structures were determined for the Rieske-type monooxygenase, stachydrine demethylase, in the unliganded state (at 1.6 Å resolution) and in the product complex (at 2.2 Å resolution). The ligand complex was obtained from enzyme aerobically cocrystallized with the substrate stachydrine (N,N-dimethylproline). The ligand electron density in the complex was interpreted as proline, generated within the active site at 100 K by the absorption of X-ray photon energy and two consecutive demethylation cycles. The oxidation state of the Rieske iron-sulfur cluster was characterized by UV-visible spectroscopy throughout X-ray data collection in conjunction with resonance Raman spectra collected before and after diffraction data. Shifts in the absorption band wavelength and intensity as a function of absorbed X-ray dose demonstrated that the Rieske center was reduced by solvated electrons generated by X-ray photons; the kinetics of the reduction process differed dramatically for the liganded complex compared to unliganded demethylase, which may correspond to the observed turnover in the crystal.

PubMed: 22224443
DOI: 10.1021/ja2111898

実験手法

X-RAY DIFFRACTION (2.2 Å)