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Accession Number ADA582327
Title Relieving Mipafox Inhibition in Organophosphorus Acid Anhydrolase by Rational Design.
Publication Date Mar 2013
Media Count 20p
Personal Author S. P. Harvey S. S. Shah T. J. Henderson
Abstract Organophosphate acid anhydrolase (OPAA) is a bimetalloenzyme that hydrolyzes acetylcholinesterase-inhibiting organophosphorus compounds, including fluorine-containing nerve agents such as soman pinacolyl methylphosphonoflouridate. The insecticide mipafox (N,N'- diisopropyldiamido fluorophosphate), which is a close analog of the nerve agent substrate diisopropyl fluorophosphonate, is a special case for OPAA because it reversibly inhibits the enzyme's hydrolysis activity. It is believed that OPAA hydrolyzes mipafox to DDP (N,N'-diisopropyld iamidophosphate), which becomes bound to the OPAA active site through a hydrogen bond network. Evaluation of the crystal structure of the OPAA-DDP complex suggests that disruption of two of these bonds, one involving Asp244 and the other involving Glu381, could potentially release DDP from the OPAA active site to give the enzyme catalytic mipafox hydrolysis activity. To test this hypothesis, we produced three mutant OPAA enzymes including one enzyme with an Asp244-to-alanine mutation, a second with a Glu381-to-alanine mutation, and a third with both mutations, to remove the corresponding hydrogen bonds between these amino acids and the bound DDP. In all cases, the mutant enzymes did not have mipafox hydrolysis activity but retained their high hydrolysis rates against soman, which indicated that the remaining OPAA-DDP interactions were adequate to stabilize their complex and keep DPP bound to the enzyme.
Keywords Acetylcholinesterase inhibition
Alteromonas bacteria
Amino acids
Bonding
Catalysts
Crystal structure
Detoxification
Enzymes
Hydrogen bonds
Hydrolysis
Inhibition
Inhibitors
Insecticides
Mipafox
Mutations
Nerve agents
Organic phosphorus compounds
Organophosphates
Organophosphorus acid hydrolyase
Substrates


 
Source Agency Non Paid ADAS
NTIS Subject Category 57B - Biochemistry
74D - Chemical, Biological, & Radiological Warfare
Corporate Author Edgewood Chemical Biological Center, Aberdeen Proving Ground, MD. Research and Technology Directorate.
Document Type Technical report
Title Note Final rept. May-Oct 2012.
NTIS Issue Number 1326
Contract Number N/A

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