Structural Insights into the Dual Activities of the Nerve Agent Degrading Organophosphate Anhydrolase/Prolidase

Abstract

The organophosphate acid anhydrolase (OPAA) is a member of a class of bimetalloenzymes that hydrolyze a variety of toxic acetylcholinesterase-inhibiting organophosphorus compounds, including fluorinecontaining chemical nerve agents. It also belongs to a family of prolidases, with significant activity against various Xaa-Pro dipeptides. Here we report the X-ray structure determination of the native OPAA (58 kDa mass) from Alteromonas sp. strain JD6.5 and its cocrystal with the inhibitor mipafox [N,N0-diisopropyldiamidofluorophosphate (DDFP)], a close analogue of the nerve agent organophosphate substrate diisopropyl fluorophosphate (DFP). The OPAA structure is composed of two domains, amino and carboxy domains, with the latter exhibiting a "pita bread" architecture and harboring the active site with the binuclear Mn2+ ions. The native OPAA structure revealed unexpectedly the presence of a well-defined nonproteinaceous density in the active site whose identity could not be definitively established but is suggestive of a bound glycolate, which is isosteric with a glycine (Xaa) product. All three glycolate oxygens coordinate the two Mn2+ atoms. DDFP or more likely its hydrolysis product, N,N0-diisopropyldiamidophosphate (DDP), is present in the cocrystal structure and bound by coordinating the binuclear metals and forming hydrogen bonds and nonpolar interactions with active site residues. An unusual common feature of the binding of the two ligands is the involvement of only one oxygen atom of the glycolate carboxylate and the product DDP tetrahedral phosphate in bridging the two Mn2 ions. Both structures provide new understanding of ligand recognition and the prolidase and organophosphorus hydrolase catalytic activities of OPAA.

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Document Details

Document Type
Technical Report
Publication Date
Dec 11, 2009
Accession Number
ADA512269

Entities

People

  • Alexei Nickitenko
  • Florante A. Quiocho
  • Nand K. Vyas
  • Saumil S. Shah
  • Vipin K. Rastogi

Organizations

  • Edgewood Chemical Biological Center

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Acetylcholinesterases
  • Amino Acids
  • Central Nervous System
  • Chemical Warfare
  • Chemical Warfare Agents
  • Chemistry
  • Crystal Structure
  • Dispersions
  • Electron Density
  • Hydrogen
  • Hydrogen Bonds
  • Nerve Agents
  • Nervous System
  • Neutral Amino Acids
  • Organophosphorus Compounds
  • Phosphorus
  • Recognition

Fields of Study

  • Chemistry

Readers

  • Molecular and Cellular Biochemistry
  • Neurotoxicology
  • Organic Chemistry