1E8H

STRUCTURE OF THE H61T MUTANT OF THE FLAVOENZYME VANILLYL-ALCOHOL OXIDASE IN THE APO FORM COMPLEXED BY ADP

1E8H の概要

• エントリーDOI: 10.2210/pdb1e8h/pdb
• 関連するPDBエントリー: 1DZN 1E0Y 1E8F 1E8G 1QLT 1QLU
• 分子名称: VANILLYL-ALCOHOL OXIDASE, ADENOSINE-5'-DIPHOSPHATE (3 entities in total)
• 機能のキーワード: oxidoreductase, flavoprotein, methanol utilization, peroxisome, flavoenzyme, oxidase, catalysis
• 由来する生物種: PENICILLIUM SIMPLICISSIMUM
• 細胞内の位置: Peroxisome: P56216
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 126794.27

構造登録者

Mattevi, A.,Fraaije, M.W. (登録日: 2000-09-20, 公開日: 2000-09-21, 最終更新日: 2023-12-13)

主引用文献

Fraaije, M.W.,Van Der Heuvel, R.H.H.,Van Berkel, W.J.H.,Mattevi, A.
Structural Analysis of Flavinylation in Vanillyl-Alcohol Oxidase
J.Biol.Chem., 275:38654-, 2001

PubMed Abstract: Vanillyl-alcohol oxidase (VAO) is member of a newly recognized flavoprotein family of structurally related oxidoreductases. The enzyme contains a covalently linked FAD cofactor. To study the mechanism of flavinylation we have created a design point mutation (His-61 --> Thr). In the mutant enzyme the covalent His-C8alpha-flavin linkage is not formed, while the enzyme is still able to bind FAD and perform catalysis. The H61T mutant displays a similar affinity for FAD and ADP (K(d) = 1.8 and 2.1 microm, respectively) but does not interact with FMN. H61T is about 10-fold less active with 4-(methoxymethyl)phenol) (k(cat) = 0.24 s(-)(1), K(m) = 40 microm) than the wild-type enzyme. The crystal structures of both the holo and apo form of H61T are highly similar to the structure of wild-type VAO, indicating that binding of FAD to the apoprotein does not require major structural rearrangements. These results show that covalent flavinylation is an autocatalytical process in which His-61 plays a crucial role by activating His-422. Furthermore, our studies clearly demonstrate that in VAO, the FAD binds via a typical lock-and-key approach to a preorganized binding site.

PubMed: 10984479
DOI: 10.1074/JBC.M004753200

実験手法

X-RAY DIFFRACTION (2.6 Å)