engleski

Acetylcholinesterase: emergence from a vulnerable target to a template for antidote and detection development

Applications of recombinant technology, chemical synthesis on biological templates and high throughput detection provide unexplored avenues for development of antidotes and arenas for remote detection for exposure to organophosphate nerve agents and pesticides. Our research strategy is based on modifying acetylcholinesterase (AChE), the very target of toxicity, so it serves in antidotal therapy and as a remote detection sensor. We discuss here how acetylcholinesterase, 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 oximes become 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 foe catalyzing the hydrolysis of organophosphates in plasma prior to their reaching cellular target site. In turn, AChE, when conjugated with organophosphate, can be employed as a template for "click chemistry" synthesis of new nucleophilic reactivating agents that could potentially prove useful in reactivation at the target site as well as in catalytic scavenging. Finally, substituted acetylcholinesterase molecules can be conjugated to fluorophores giving rise to shifts in emission spectra for detection of exposure to organophosphates. Since reagents do not have to be added to detect the fluorescence change, the modified enzyme would serve as a remota sensor. Supported by UO1-ES010337 and R37-GM18360 from the National Institutes of Health.

acetylcholinesterase; mutants; oximes; click chemistry; remote sensors

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