title: Bioactivation of chlorpyrifos by CYP2B6 variants.
abstract: Chlorpyrifos (CPF), an organophosphorus (OP) pesticide, is bioactivated by cytochrome P450s (CYPs) to the active metabolite chlorpyrifos oxon (CPF-O). Given that human CYP2B6 has the highest intrinsic clearance (CL(int)) for CPF bioactivation, CYP2B6 polymorphisms may impact human susceptibility to CPF at real world environmental and occupational CPF exposure levels. CYP2B6.4,.5,.7, and .18 were over-expressed in mammalian COS-1 cells to assess the impact of CYP2B6 variants on the K(m) and V(max) for bioactivation of CPF. Cell lysates were incubated with CPF (0-100 μM) and the production of CPF-O was measured via HPLC analysis. CYP2B6 content was determined by western blot. CYP2B6.18 had neither detectable protein nor activity levels. The V(max) value for each remaining variant was significantly higher than wild-type (CYP2B6.1, V(max) 4.13 × 10(4) pmol/min/nmol CYP2B6), with CYP2B6.4,.5, and .7 having V(max) values of 4.52 × 10(5), 1.82 × 10(5), and 9.60 × 10(4) pmol/min/nmol CYP2B6, respectively. The K(m) values for these variants ranged from 0.39 to 1.09 μM and were not significantly different from wild-type. All active variants examined had significantly higher CL(int) than CYP2B6.1. Variants of CYP2B6 have altered capacity to bioactivate CPF and may affect individual susceptibility by altering the V(max) for CPF-O formation.
authors: Department of Pharmacology and Toxicology, State University of New York at Buffalo, Buffalo, NY, USA.
authors: Alice L
authors: AL
authors: Crane
authors: Kathrin
authors: K
authors: Klein
authors: James R
authors: JR
authors: Olson
mesh_terms: D001741:Black or African American / Q000235:genetics; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics* / Q000378:metabolism*; D001711:Biotransformation; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D004390:Chlorpyrifos / Q000737:chemistry / Q000378:metabolism*; D065702:Cytochrome P-450 CYP2B6; D006801:Humans; D007700:Kinetics; D053858:Metabolic Networks and Pathways; D008862:Microsomes, Liver / Q000201:enzymology; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics* / Q000378:metabolism*; D011110:Polymorphism, Genetic; D013379:Substrate Specificity; D044465:White People / Q000235:genetics
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; C585599:CYP2B6 protein, human; D065702:Cytochrome P-450 CYP2B6; D010089:Oxidoreductases, N-Demethylating; D004390:Chlorpyrifos