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dc.contributor.authorBester, S.M.
dc.contributor.authorAdipietro, K.A.
dc.contributor.authorFunk, V.L.
dc.date.accessioned2019-09-13T17:02:32Z
dc.date.available2019-09-13T17:02:32Z
dc.date.issued2019
dc.identifier.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85065438023&doi=10.1002%2fpro.3625&partnerID=40&md5=36693c2c6f993501b831694f57d94f92
dc.identifier.urihttp://hdl.handle.net/10713/10798
dc.description.abstractServing a critical role in neurotransmission, human acetylcholinesterase (hAChE) is the target of organophosphate nerve agents. Hence, there is an active interest in studying the mechanism of inhibition and recovery of enzymatic activity, which could lead to better countermeasures against nerve agents. As hAChE is found in different oligomeric assemblies, certain approaches to studying it have been problematic. Herein, we examine the biochemical and structural impact of monomerizing hAChE by using two mutations: L380R/F535K. The activities of monomeric hAChE L380R/F535K and dimeric hAChE were determined to be comparable utilizing a modified Ellman's assay. To investigate the influence of subunit–subunit interactions on the structure of hAChE, a 2.1 Å X‐ray crystallographic structure was determined. Apart from minor shifts along the dimer interface, the overall structure of the hAChE L380R/F535K mutant is similar to that of dimeric hAChE. To probe whether the plasticity of the active site was overtly impacted by monomerizing hAChE, the kinetic constants of (PR/S) − VX (ethyl({2‐[bis(propan‐2‐yl)amino]ethyl}sulfanyl)(methyl)phosphinate) inhibition and subsequent rescue of hAChE L380R/F535K activity with HI‐6 (1‐(2′‐hydroxyiminomethyl‐1′‐pyridinium)‐3‐(4′‐carbamoyl‐1‐pyridinium)) were determined and found to be comparable to those of dimeric hAChE. Thus, hAChE L380R/F535K could be used as a substitute for dimeric hAChE when experimentally probing the ability of the hAChE active site to accommodate future nerve agent threats or judge the ability of new therapeutics to access the active site. Copyright 2019 The Authors.en_US
dc.description.sponsorshipThis work was funded by the Defense Threat Reduction Agency project: CB#3889 ?Elucidation of the mechanisms and physical properties of the molecular targets of chemical nerve agents? (JJH & SDP).en_US
dc.description.urihttps://doi.org/10.1002/pro.3625en_US
dc.language.isoen-USen_US
dc.publisherBlackwell Publishing Ltden_US
dc.relation.ispartofProtein Science
dc.subjectacetylcholinesteraseen_US
dc.subjectnerve agenten_US
dc.subjectoligomerizationen_US
dc.subjectorganophosphateen_US
dc.titleThe structural and biochemical impacts of monomerizing human acetylcholinesteraseen_US
dc.typeArticleen_US
dc.identifier.doi10.1002/pro.3625
dc.identifier.pmid30993792


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