title: Human carboxylesterase 1 stereoselectively binds the nerve agent cyclosarin and spontaneously hydrolyzes the nerve agent sarin.
abstract: Organophosphorus (OP) nerve agents are potent toxins that inhibit cholinesterases and produce a rapid and lethal cholinergic crisis. Development of protein-based therapeutics is being pursued with the goal of preventing nerve agent toxicity and protecting against the long-term side effects of these agents. The drug-metabolizing enzyme human carboxylesterase 1 (hCE1) is a candidate protein-based therapeutic because of its similarity in structure and function to the cholinesterase targets of nerve agent poisoning. However, the ability of wild-type hCE1 to process the G-type nerve agents sarin and cyclosarin has not been determined. We report the crystal structure of hCE1 in complex with the nerve agent cyclosarin. We further use stereoselective nerve agent analogs to establish that hCE1 exhibits a 1700- and 2900-fold preference for the P(R) enantiomers of analogs of soman and cyclosarin, respectively, and a 5-fold preference for the P(S) isomer of a sarin analog. Finally, we show that for enzyme inhibited by racemic mixtures of bona fide nerve agents, hCE1 spontaneously reactivates in the presence of sarin but not soman or cyclosarin. The addition of the neutral oxime 2,3-butanedione monoxime increases the rate of reactivation of hCE1 from sarin inhibition by more than 60-fold but has no effect on reactivation with the other agents examined. Taken together, these data demonstrate that hCE1 is only reactivated after inhibition with the more toxic P(S) isomer of sarin. These results provide important insights toward the long-term goal of designing novel forms of hCE1 to act as protein-based therapeutics for nerve agent detoxification.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3290, USA.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Hemmert
authors.forename: Tamara C
authors.initials: TC
authors.lastname: Otto
authors.forename: Monika
authors.initials: M
authors.lastname: Wierdl
authors.forename: Carol C
authors.initials: CC
authors.lastname: Edwards
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Fleming
authors.forename: Mary
authors.initials: M
authors.lastname: MacDonald
authors.forename: John R
authors.initials: JR
authors.lastname: Cashman
authors.forename: Philip M
authors.initials: PM
authors.lastname: Potter
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Cerasoli
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Redinbo
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D002619:Chemical Warfare Agents / Q000737:chemistry*; D003460:Crystallization; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D008958:Models, Molecular; D009943:Organophosphorus Compounds / Q000737:chemistry* / Q000494:pharmacology; D010091:Oximes / Q000494:pharmacology; D012524:Sarin / Q000737:chemistry* / Q000494:pharmacology; D013237:Stereoisomerism
chemical_list: D002619:Chemical Warfare Agents; D004791:Enzyme Inhibitors; D009943:Organophosphorus Compounds; D010091:Oximes; D012524:Sarin; D002265:Carboxylic Ester Hydrolases; C474609:CES1 protein, human; C053914:cyclohexyl methylphosphonofluoridate