2WOH

Strontium soaked E. coli copper amine oxidase

2WOH の概要

• エントリーDOI: 10.2210/pdb2woh/pdb
• 関連するPDBエントリー: 1D6U 1D6Y 1D6Z 1DYU 1JRQ 1LVN 1OAC 1QAF 1QAK 1QAL 1SPU 2W0Q 2WGQ 2WO0 2WOF
• 分子名称: PRIMARY AMINE OXIDASE, COPPER (II) ION, CALCIUM ION, ... (5 entities in total)
• 機能のキーワード: tpq, metal-binding, oxidoreductase, amine oxidation
• 由来する生物種: ESCHERICHIA COLI
• 細胞内の位置: Periplasm: P46883
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 163115.94

構造登録者

Smith, M.A.,Pirrat, P.,Pearson, A.R.,Knowles, P.F.,Phillips, S.E.V.,McPherson, M.J. (登録日: 2009-07-23, 公開日: 2010-05-05, 最終更新日: 2025-04-09)

主引用文献

Smith, M.A.,Pirrat, P.,Pearson, A.R.,Kurtis, C.R.,Gaule, T.G.,Knowles, P.F.,Phillips, S.E.,McPherson, M.J.
Exploring the Roles of the Metal Ions in Escherichia Coli Copper Amine Oxidase.
Biochemistry, 49:1268-, 2010

PubMed Abstract: To investigate the role of the active site copper in Escherichia coli copper amine oxidase (ECAO), we initiated a metal-substitution study. Copper reconstitution of ECAO (Cu-ECAO) restored only approximately 12% wild-type activity as measured by k(cat(amine)). Treatment with EDTA, to remove exogenous divalent metals, increased Cu-ECAO activity but reduced the activity of wild-type ECAO. Subsequent addition of calcium restored wild-type ECAO and further enhanced Cu-ECAO activities. Cobalt-reconstituted ECAO (Co-ECAO) showed lower but significant activity. These initial results are consistent with a direct electron transfer from TPQ to oxygen stabilized by the metal. If a Cu(I)-TPQ semiquinone mechanism operates, then an alternative outer-sphere electron transfer must also exist to account for the catalytic activity of Co-ECAO. The positive effect of calcium on ECAO activity led us to investigate the peripheral calcium binding sites of ECAO. Crystallographic analysis of wild-type ECAO structures, determined in the presence and absence of EDTA, confirmed that calcium is the normal ligand of these peripheral sites. The more solvent exposed calcium can be easily displaced by mono- and divalent cations with no effect on activity, whereas removal of the more buried calcium ion with EDTA resulted in a 60-90% reduction in ECAO activity and the presence of a lag phase, which could be overcome under oxygen saturation or by reoccupying the buried site with various divalent cations. Our studies indicate that binding of metal ions in the peripheral sites, while not essential, is important for maximal enzymatic activity in the mature enzyme.

PubMed: 20052994
DOI: 10.1021/BI901738K

実験手法

X-RAY DIFFRACTION (2.7 Å)