4ZWO

Crystal structure of organophosphate anhydrolase/prolidase mutant Y212F

Summary for 4ZWO

• Entry DOI: 10.2210/pdb4zwo/pdb
• Related: 3L24 3L7G 4ZWP 4ZWU
• Descriptor: organophosphate anhydrolase/prolidase, MANGANESE (II) ION, GLYCOLIC ACID, ... (5 entities in total)
• Functional Keywords: opaa organophosphate prolidase anhydrolase, hydrolase
• Biological source: Alteromonas sp.
• Total number of polymer chains: 2
• Total formula weight: 101407.92

Authors

Daczkowski, C.M.,Pegan, S.D.,Harvey, S.P. (deposition date: 2015-05-19, release date: 2015-10-14, Last modification date: 2023-11-15)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.141 Å)