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dc.contributor.authorTemeyer, Kevin Ben_US
dc.contributor.authorTong, Fanen_US
dc.contributor.authorTotrov, Maxim Men_US
dc.contributor.authorTuckow, Alexander Pen_US
dc.contributor.authorChen, Qiao-hongen_US
dc.contributor.authorCarlier, Paul Ren_US
dc.contributor.authorPérez de León, Adalberto Aen_US
dc.contributor.authorBloomquist, Jeffrey Ren_US
dc.date.accessioned2014-12-16T16:15:15Z
dc.date.available2014-12-16T16:15:15Z
dc.date.issued2014-12-10
dc.identifier.citationParasites & Vectors. 2014 Dec 10;7(1):577en_US
dc.identifier.urihttp://hdl.handle.net/10919/51141
dc.description.abstractBackground Phlebotomus papatasi vectors zoonotic cutaneous leishmaniasis. Previous expression of recombinant P. papatasi acetylcholinesterase (PpAChE1) revealed 85% amino acid sequence identity to mosquito AChE and identified synthetic carbamates that effectively inhibited PpAChE1 with improved specificity for arthropod AChEs compared to mammalian AChEs. We hypothesized that the G119S mutation causing high level resistance to organophosphate insecticides in mosquitoes may occur in PpAChE1 and may reduce sensitivity to inhibition. We report construction, expression, and biochemical properties of rPpAChE1 containing the G119S orthologous mutation. Methods Targeted mutagenesis introduced the G119S orthologous substitution in PpAChE1 cDNA. Recombinant PpAChE1 enzymes containing or lacking the G119S mutation were expressed in the baculoviral system. Biochemical assays were conducted to determine altered catalytic properties and inhibitor sensitivity resulting from the G119S substitution. A molecular homology model was constructed to examine the modeled structural interference with docking of inhibitors of different classes. Genetic tests were conducted to determine if the G119S orthologous codon existed in polymorphic form in a laboratory colony of P. papatasi. Results Recombinant PpAChE1 containing the G119S substitution exhibited altered biochemical properties, and reduced inhibition by compounds that bind to the acylation site on the enzyme (with the exception of eserine). Less resistance was directed against bivalent or peripheral site inhibitors, in good agreement with modeled inhibitor docking. Eserine appeared to be a special case capable of inhibition in the absence of covalent binding at the acylation site. Genetic tests did not detect the G119S mutation in a laboratory colony of P. papatasi but did reveal that the G119S codon existed in polymorphic form (GGA + GGC). Conclusions The finding of G119S codon polymorphism in a laboratory colony of P. papatasi suggests that a single nucleotide transversion (GGC → AGC) may readily occur, causing rapid development of resistance to organophosphate and phenyl-substituted carbamate insecticides under strong selection. Careful management of pesticide use in IPM programs is important to prevent or mitigate development and fixation of the G119S mutation in susceptible pest populations. Availability of recombinant AChEs enables identification of novel inhibitory ligands with improved efficacy and specificity for AChEs of arthropod pests.en_US
dc.format.mimetypeapplication/pdf
dc.language.isoen_US
dc.rightsCreative Commons Attribution 4.0 International*
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/*
dc.titleAcetylcholinesterase of the sand fly, Phlebotomus papatasi (Scopoli): construction, expression and biochemical properties of the G119S orthologous mutanten_US
dc.typeArticle - Refereed
dc.date.updated2014-12-16T16:15:15Z
dc.description.versionPeer Reviewed
dc.rights.holderKevin B Temeyer et al.; licensee BioMed Central Ltd.en_US
dc.title.serialParasites & Vectors
dc.identifier.doihttps://doi.org/10.1186/s13071-014-0577-4
dc.type.dcmitypeText


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Creative Commons Attribution 4.0 International
License: Creative Commons Attribution 4.0 International