title: Interaction of the serine hydrolase KIAA1363 with organophosphorus agents: Evaluation of potency and kinetics.
abstract: Oxons are bioactive metabolites of organophosphorus insecticides (OPs) that covalently inactivate serine hydrolases. KIAA1363 is one of the most abundant serine hydrolases in mouse brain. Although the physiological consequences related to the inhibition of KIAA1363 due to environmental exposures to OPs are poorly understood, the enzyme was previously shown to have a role in the detoxification of oxons. Here, we overexpressed human KIAA1363 and CES1 in COS7 cells and compared the potency of inhibition (IC50s, 15 min) of KIAA1363 and CES1 by chlorpyrifos oxon (CPO), paraoxon (PO), and methyl paraoxon (MPO). The order of potency was CPO > PO >> MPO for both enzymes. We also determined the bimolecular rate constants (kinact/Ki) for reactions of CPO and PO with KIAA1363 and CES1. KIAA1363 and CES1 were inactivated by CPO at comparable rates (4.4 × 10(6) s(-1) M(-1) and 6.7 × 10(6) s(-1) M(-1), respectively), whereas PO inactivated both enzymes at slower rates (0.4 × 10(6) s(-1) M(-1) and 1.5 × 10(6) s(-1) M(-1), respectively). Finally, the reactivation rate of KIAA1363 following inhibition by CPO was evaluated. Together, the results define the kinetics of inhibition of KIAA1363 by active metabolites of agrochemicals and indicate that KIAA1363 is highly sensitive to inhibition by these compounds.
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States; Center for Environmental Health Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States. Electronic address: mross@cvm.msstate.edu.
authors.forename: Matthew K
authors.initials: MK
authors.lastname: Ross
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States.
authors.forename: Kim
authors.initials: K
authors.lastname: Pluta
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States.
authors.forename: Victoria
authors.initials: V
authors.lastname: Bittles
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States; Center for Environmental Health Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States.
authors.forename: Abdolsamad
authors.initials: A
authors.lastname: Borazjani
authors.affiliation: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States; Center for Environmental Health Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, United States. Electronic address: crow@cvm.msstate.edu.
authors.forename: J
authors.initials: J
authors.lastname: Allen Crow
mesh_terms: D000818:Animals; D019556:COS Cells; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D002522:Chlorocebus aethiops; D002801:Cholinesterase Reactivators / Q000737:chemistry* / Q000378:metabolism*; D004789:Enzyme Activation; D007700:Kinetics; D008657:Metabolic Clearance Rate; D008954:Models, Biological; D008956:Models, Chemical; D010755:Organophosphates / Q000737:chemistry* / Q000378:metabolism*; D002787:Sterol Esterase; D013379:Substrate Specificity
chemical_list: D002801:Cholinesterase Reactivators; D010755:Organophosphates; D002265:Carboxylic Ester Hydrolases; C515677:NCEH1 protein, human; D002787:Sterol Esterase
keywords: Bimolecular rate constant; Carboxylesterase; Chlorpyrifos oxon; Enzyme inhibition; Oxon; Paraoxon