3CMC

Thioacylenzyme intermediate of Bacillus stearothermophilus phosphorylating GAPDH

3CMC の概要

• エントリーDOI: 10.2210/pdb3cmc/pdb
• 関連するPDBエントリー: 1GD1 1NPT 1NQ5 1NQA 1NQO 2GD1
• 分子名称: Glyceraldehyde-3-phosphate dehydrogenase, SULFATE ION, GLYCERALDEHYDE-3-PHOSPHATE, ... (7 entities in total)
• 機能のキーワード: microspectrophotometry, reaction intermediate, dehydrogenase, phosphate binding site, thioacylenzyme, glycolysis, nad, oxidoreductase
• 由来する生物種: Bacillus stearothermophilus
• 細胞内の位置: Cytoplasm: P00362
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 149826.07

構造登録者

Moniot, S.,Vonrhein, C.,Bricogne, G.,Didierjean, C.,Corbier, C. (登録日: 2008-03-21, 公開日: 2008-06-17, 最終更新日: 2024-10-30)

主引用文献

Moniot, S.,Bruno, S.,Vonrhein, C.,Didierjean, C.,Boschi-Muller, S.,Vas, M.,Bricogne, G.,Branlant, G.,Mozzarelli, A.,Corbier, C.
Trapping of the Thioacylglyceraldehyde-3-phosphate Dehydrogenase Intermediate from Bacillus stearothermophilus: DIRECT EVIDENCE FOR A FLIP-FLOP MECHANISM
J.Biol.Chem., 283:21693-21702, 2008

PubMed Abstract: The crystal structure of the thioacylenzyme intermediate of the phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus has been solved at 1.8A resolution. Formation of the intermediate was obtained by diffusion of the natural substrate within the crystal of the holoenzyme in the absence of inorganic phosphate. To define the soaking conditions suitable for the isolation and accumulation of the intermediate, a microspectrophotometric characterization of the reaction of GAPDH in single crystals was carried out, following NADH formation at 340 nm. When compared with the structure of the Michaelis complex (Didierjean, C., Corbier, C., Fatih, M., Favier, F., Boschi-Muller, S., Branlant, G., and Aubry, A. (2003) J. Biol. Chem. 278, 12968-12976) the 206-210 loop is shifted and now forms part of the so-called "new P(i)" site. The locations of both the O1 atom and the C3-phosphate group of the substrate are also changed. Altogether, the results provide evidence for the flipping of the C3-phosphate group occurring concomitantly or after the redox step.

PubMed: 18480053
DOI: 10.1074/jbc.M802286200

実験手法

X-RAY DIFFRACTION (1.77 Å)