3E2N

Engineering ascorbate peroxidase activity into cytochrome c peroxidase

3E2N の概要

• エントリーDOI: 10.2210/pdb3e2n/pdb
• 関連するPDBエントリー: 1DSG 1DSO 1ZBY 2V23 3E2O 5CCP
• 分子名称: Cytochrome c peroxidase, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• 機能のキーワード: cytochrome c peroxidase (ccp), ascorbate peroxidase (apx), oxidoreductase
• 由来する生物種: Saccharomyces cerevisiae; 詳細
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 33218.96

構造登録者

Poulos, T.L.,Meharenna, Y.T.,Oertel, P. (登録日: 2008-08-05, 公開日: 2008-10-21, 最終更新日: 2023-08-30)

主引用文献

Meharenna, Y.T.,Oertel, P.,Bhaskar, B.,Poulos, T.L.
Engineering ascorbate peroxidase activity into cytochrome c peroxidase.
Biochemistry, 47:10324-10332, 2008

PubMed Abstract: Cytochrome c peroxidase (CCP) and ascorbate peroxidase (APX) have very similar structures, and yet neither CCP nor APX exhibits each other's activities with respect to reducing substrates. APX has a unique substrate binding site near the heme propionates where ascorbate H-bonds with a surface Arg and one heme propionate (Sharp et al. (2003) Nat. Struct. Biol. 10, 303-307). The corresponding region in CCP has a much longer surface loop, and the critical Arg residue that is required for ascorbate binding in APX is Asn in CCP. In order to convert CCP into an APX, the ascorbate-binding loop and critical arginine were engineered into CCP to give the CCP2APX mutant. The mutant crystal structure shows that the engineered site is nearly identical to that found in APX. While wild-type CCP shows no APX activity, CCP2APX catalyzes the peroxidation of ascorbate at a rate of approximately 12 min (-1), indicating that the engineered ascorbate-binding loop can bind ascorbate.

PubMed: 18771292
DOI: 10.1021/bi8007565

実験手法

X-RAY DIFFRACTION (1.3 Å)