title: Acetylcholinesterase of Stomoxys calcitrans (L.) (Diptera: Muscidae): cDNA sequence, baculovirus expression, and biochemical properties.
abstract: A 2193-nucleotide cDNA encoding acetylcholinesterase (AChE) of the stable fly, Stomoxys calcitrans (L.) was sequenced and expressed in the baculovirus system. The open reading frame encoded a 91 amino acid secretion signal peptide and a 613 amino acid mature protein with 96% and 94% identity to the AChEs of Haematobia irritans (L.) and Musca domestica (L.), respectively. Structural characteristics of M. domestica, H. irritans, and Drosophila melanogaster AChEs were conserved in the S. calcitrans AChE. The recombinant enzyme was inhibited by eserine, coroxon, and paraoxon and exhibited K(m) values of 63.9μM for acetylthiocholine and 96.7μM for butyrylthiocholine, confirming its biochemical identity as an acetylcholinesterase (EC 3.1.1.7). These data will enable rapid identification and assay for mutations that reduce AChE sensitivity to organophosphate (OP) pesticides, potentially aiding resistance management efforts to prevent fixation of the mutations in pest populations.
authors.affiliation: Knipling-Bushland U.S. Livestock Insects Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 2700 Fredericksburg Road, Kerrville, TX 78028-9184, USA. Kevin.Temeyer@ars.usda.gov
authors.forename: Kevin B
authors.initials: KB
authors.lastname: Temeyer
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Chen
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008969:Molecular Sequence Data; D009117:Muscidae / Q000201:enzymology* / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D000110:Acetylcholinesterase