4ZWU

Crystal structure of organophosphate anhydrolase/prolidase mutant Y212F, V342L, I215Y

4ZWU の概要

• エントリーDOI: 10.2210/pdb4zwu/pdb
• 関連するPDBエントリー: 4ZWO 4ZWP
• 分子名称: ORGANOPHOSPHATE ANHYDROLASE/PROLIDASE, MANGANESE (II) ION, BARIUM ION, ... (6 entities in total)
• 機能のキーワード: opaa organophosphate prolidase anhydrolase, hydrolase
• 由来する生物種: Alteromonas sp.
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 101679.67

構造登録者

Daczkowski, C.M.,Pegan, S.D.,Harvey, S.P. (登録日: 2015-05-19, 公開日: 2015-10-14, 最終更新日: 2023-11-15)

主引用文献

Daczkowski, C.M.,Pegan, S.D.,Harvey, S.P.
Engineering the Organophosphorus Acid Anhydrolase Enzyme for Increased Catalytic Efficiency and Broadened Stereospecificity on Russian VX.
Biochemistry, 54:6423-6433, 2015

PubMed Abstract: The enzyme organophosphorus acid anhydrolase (OPAA), from Alteromonas sp. JD6.5, has been shown to rapidly catalyze the hydrolysis of a number of toxic organophosphorus compounds, including several G-type chemical nerve agents. The enzyme was cloned into Escherichia coli and can be produced up to approximately 50% of cellular protein. There have been no previous reports of OPAA activity on VR {Russian VX, O-isobutyl S-[2-(diethylamino)ethyl] methylphosphonothioate}, and our studies reported here show that wild-type OPAA has poor catalytic efficacy toward VR. However, via application of a structurally aided protein engineering approach, significant improvements in catalytic efficiency were realized via optimization of the small pocket within the OPAA's substrate-binding site. This optimization involved alterations at only three amino acid sites resulting in a 30-fold increase in catalytic efficiency toward racemic VR, with a strong stereospecificity toward the P(+) enantiomer. X-ray structures of this mutant as well as one of its predecessors provide potential structural rationales for their effect on the OPAA active site. Additionally, a fourth mutation at a site near the small pocket was found to relax the stereospecificity of the OPAA enzyme. Thus, it allows the altered enzyme to effectively process both VR enantiomers and should be a useful genetic background in which to seek further improvements in OPAA VR activity.

PubMed: 26418828
DOI: 10.1021/acs.biochem.5b00624

実験手法

X-RAY DIFFRACTION (2.2 Å)