3T1G

Engineering of organophosphate hydrolase by computational design and directed evolution

3T1G の概要

• エントリーDOI: 10.2210/pdb3t1g/pdb
• 関連するPDBエントリー: 1A4L
• 分子名称: organophosphate hydrolase, ZINC ION, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: computational design, directed evolution, tim beta/alpha-barrel, metallo-dependent hydrolase, organophosphate binding, hydrolysis, artificial enzyme, hydrolase
• 由来する生物種: Mus musculus (mouse)
• 細胞内の位置: Cell membrane; Peripheral membrane protein; Extracellular side (By similarity): P03958
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 40482.39

構造登録者

Takeuchi, R.,Stoddard, B.L. (登録日: 2011-07-21, 公開日: 2012-02-08, 最終更新日: 2023-09-13)

主引用文献

Khare, S.D.,Kipnis, Y.,Greisen, P.J.,Takeuchi, R.,Ashani, Y.,Goldsmith, M.,Song, Y.,Gallaher, J.L.,Silman, I.,Leader, H.,Sussman, J.L.,Stoddard, B.L.,Tawfik, D.S.,Baker, D.
Computational redesign of a mononuclear zinc metalloenzyme for organophosphate hydrolysis.
Nat.Chem.Biol., 8:294-300, 2012

PubMed Abstract: The ability to redesign enzymes to catalyze noncognate chemical transformations would have wide-ranging applications. We developed a computational method for repurposing the reactivity of metalloenzyme active site functional groups to catalyze new reactions. Using this method, we engineered a zinc-containing mouse adenosine deaminase to catalyze the hydrolysis of a model organophosphate with a catalytic efficiency (k(cat)/K(m)) of ~10(4) M(-1) s(-1) after directed evolution. In the high-resolution crystal structure of the enzyme, all but one of the designed residues adopt the designed conformation. The designed enzyme efficiently catalyzes the hydrolysis of the R(P) isomer of a coumarinyl analog of the nerve agent cyclosarin, and it shows marked substrate selectivity for coumarinyl leaving groups. Computational redesign of native enzyme active sites complements directed evolution methods and offers a general approach for exploring their untapped catalytic potential for new reactivities.

PubMed: 22306579
DOI: 10.1038/nchembio.777

実験手法

X-RAY DIFFRACTION (2.35 Å)