Acetylcholinesterase: converting a vulnerable target to a template for antidotes and detection of inhibitor exposure.
Applications of recombinant DNA technology, chemical synthesis on biological templates and fluorescence detection of organophosphorylation provide unexplored avenues for development of antidotes and approaches for remote detection of organophosphate nerve agents and pesticides. We discuss here how acetylcholinesterase (AChE), through appropriate mutations, becomes more susceptible to oxime reactivation. Since the reaction between organophosphate and the mutated enzyme remains rapid, regeneration of active enzyme by oxime becomes the rate-limiting step in the process to complete a catalytic cycle for generation of active enzyme. Accordingly, "Oxime-assisted Catalysis" by AChE provides a potential means for catalyzing the hydrolysis of organophosphates in plasma prior to their reaching the cellular target site. In turn, AChE, when conjugated with organophosphate, is employed as a template for 'click-chemistry, freeze-frame' synthesis of new nucleophilic reactivating agents that could potentially prove useful in AChE reactivation at the target site as well as in catalytic scavenging of organophosphates in plasma. Finally, substituted AChE molecules can be conjugated to fluorophores giving rise to shifts in emission spectra for detection of dispersed organophosphates. Since external reagents do not have to be added to detect the fluorescence change, the modified enzyme would serve as a remote sensor.
Taylor
Palmer
P
Department of Pharmacology, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA 92093-0650, USA. pwtaylor@ucsd.edu
Kovarik
Zrinka
Z
Reiner
Elsa
E
Radić
Zoran
Z
D000110:Acetylcholinesterase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000931:Antidotes / Q000737:chemistry / Q000494:pharmacology; D001667:Binding, Competitive; D002384:Catalysis; D002800:Cholinesterase Inhibitors / Q000032:analysis / Q000493:pharmacokinetics / Q000506:poisoning; D002801:Cholinesterase Reactivators / Q000737:chemistry / Q000494:pharmacology; D015195:Drug Design; D006801:Humans; D006868:Hydrolysis; D008024:Ligands; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D062025:Organophosphate Poisoning; D009943:Organophosphorus Compounds / Q000032:analysis / Q000493:pharmacokinetics; D010091:Oximes / Q000737:chemistry / Q000494:pharmacology; D011041:Poisoning / Q000188:drug therapy / Q000201:enzymology; D013050:Spectrometry, Fluorescence
D000931:Antidotes; D002800:Cholinesterase Inhibitors; D002801:Cholinesterase Reactivators; D008024:Ligands; D009943:Organophosphorus Compounds; D010091:Oximes; D000110:Acetylcholinesterase