3EYA

Structural basis for membrane binding and catalytic activation of the peripheral membrane enzyme pyruvate oxidase from Escherichia coli

3EYA の概要

• エントリーDOI: 10.2210/pdb3eya/pdb
• 関連するPDBエントリー: 1POW 3EY9
• 分子名称: Pyruvate dehydrogenase [cytochrome], THIAMINE DIPHOSPHATE, FLAVIN-ADENINE DINUCLEOTIDE, ... (6 entities in total)
• 機能のキーワード: pyruvate oxidase, membrane-associated flavoprotein dehydrogenase, interactions with lipids cell membrane, fad, flavoprotein, lipid-binding, magnesium, membrane, oxidoreductase, thiamine pyrophosphate
• 由来する生物種: Escherichia coli
• 細胞内の位置: Cell membrane; Peripheral membrane protein: P07003
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 731702.48

構造登録者

Neumann, P.,Weidner, A.,Pech, A.,Stubbs, M.T.,Tittmann, K. (登録日: 2008-10-20, 公開日: 2008-11-04, 最終更新日: 2023-11-01)

主引用文献

Neumann, P.,Weidner, A.,Pech, A.,Stubbs, M.T.,Tittmann, K.
Structural basis for membrane binding and catalytic activation of the peripheral membrane enzyme pyruvate oxidase from Escherichia coli.
Proc.Natl.Acad.Sci.USA, 105:17390-17395, 2008

PubMed Abstract: The thiamin- and flavin-dependent peripheral membrane enzyme pyruvate oxidase from E. coli catalyzes the oxidative decarboxylation of the central metabolite pyruvate to CO(2) and acetate. Concomitant reduction of the enzyme-bound flavin triggers membrane binding of the C terminus and shuttling of 2 electrons to ubiquinone 8, a membrane-bound mobile carrier of the electron transport chain. Binding to the membrane in vivo or limited proteolysis in vitro stimulate the catalytic proficiency by 2 orders of magnitude. The molecular mechanisms by which membrane binding and activation are governed have remained enigmatic. Here, we present the X-ray crystal structures of the full-length enzyme and a proteolytically activated truncation variant lacking the last 23 C-terminal residues inferred as important in membrane binding. In conjunction with spectroscopic results, the structural data pinpoint a conformational rearrangement upon activation that exposes the autoinhibitory C terminus, thereby freeing the active site. In the activated enzyme, Phe-465 swings into the active site and wires both cofactors for efficient electron transfer. The isolated C terminus, which has no intrinsic helix propensity, folds into a helical structure in the presence of micelles.

PubMed: 18988747
DOI: 10.1073/pnas.0805027105

実験手法

X-RAY DIFFRACTION (2.5 Å)