2EZ9

Pyruvate oxidase variant F479W in complex with reaction intermediate analogue 2-phosphonolactyl-thiamin diphosphate

2EZ9 の概要

• エントリーDOI: 10.2210/pdb2ez9/pdb
• 関連するPDBエントリー: 1POW 1POX 1Y9D 2EZ4 2EZ8 2EZT 2EZU
• 分子名称: Pyruvate oxidase, MAGNESIUM ION, SODIUM ION, ... (6 entities in total)
• 機能のキーワード: tpp enzyme, reaction intermediate analogue, oxidoreductase
• 由来する生物種: Lactobacillus plantarum
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 135284.11

構造登録者

Wille, G.,Meyer, D.,Steinmetz, A.,Hinze, E.,Golbik, R.,Tittmann, K. (登録日: 2005-11-10, 公開日: 2006-04-25, 最終更新日: 2023-08-23)

主引用文献

Wille, G.,Meyer, D.,Steinmetz, A.,Hinze, E.,Golbik, R.,Tittmann, K.
The catalytic cycle of a thiamin diphosphate enzyme examined by cryocrystallography.
Nat.Chem.Biol., 2:324-328, 2006

PubMed Abstract: Enzymes that use the cofactor thiamin diphosphate (ThDP, 1), the biologically active form of vitamin B(1), are involved in numerous metabolic pathways in all organisms. Although a theory of the cofactor's underlying reaction mechanism has been established over the last five decades, the three-dimensional structures of most major reaction intermediates of ThDP enzymes have remained elusive. Here, we report the X-ray structures of key intermediates in the oxidative decarboxylation of pyruvate, a central reaction in carbon metabolism catalyzed by the ThDP- and flavin-dependent enzyme pyruvate oxidase (POX)3 from Lactobacillus plantarum. The structures of 2-lactyl-ThDP (LThDP, 2) and its stable phosphonate analog, of 2-hydroxyethyl-ThDP (HEThDP, 3) enamine and of 2-acetyl-ThDP (AcThDP, 4; all shown bound to the enzyme's active site) provide profound insights into the chemical mechanisms and the stereochemical course of thiamin catalysis. These snapshots also suggest a mechanism for a phosphate-linked acyl transfer coupled to electron transfer in a radical reaction of pyruvate oxidase.

PubMed: 16680160
DOI: 10.1038/nchembio788

実験手法

X-RAY DIFFRACTION (1.6 Å)