title: Pyrethroid activity-based probes for profiling cytochrome P450 activities associated with insecticide interactions.
abstract: Pyrethroid insecticides are used to control diseases spread by arthropods. We have developed a suite of pyrethroid mimetic activity-based probes (PyABPs) to selectively label and identify P450s associated with pyrethroid metabolism. The probes were screened against pyrethroid-metabolizing and nonmetabolizing mosquito P450s, as well as rodent microsomes, to measure labeling specificity, plus cytochrome P450 oxidoreductase and b5 knockout mouse livers to validate P450 activation and establish the role for b5 in probe activation. Using PyABPs, we were able to profile active enzymes in rat liver microsomes and identify pyrethroid-metabolizing enzymes in the target tissue. These included P450s as well as related detoxification enzymes, notably UDP-glucuronosyltransferases, suggesting a network of associated pyrethroid-metabolizing enzymes, or "pyrethrome." Considering the central role P450s play in metabolizing insecticides, we anticipate that PyABPs will aid in the identification and profiling of P450s associated with insecticide pharmacology in a wide range of species, improving understanding of P450-insecticide interactions and aiding the development of unique tools for disease control.
authors.affiliation: Vector Biology, Liverpool School of Tropical Medicine, Liverpool L3 5QA, United Kingdom.
authors.forename: Hanafy M
authors.initials: HM
authors.lastname: Ismail
authors.forename: Paul M
authors.initials: PM
authors.lastname: O'Neill
authors.forename: David W
authors.initials: DW
authors.lastname: Hong
authors.forename: Robert D
authors.initials: RD
authors.lastname: Finn
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Henderson
authors.forename: Aaron T
authors.initials: AT
authors.lastname: Wright
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
authors.forename: Janet
authors.initials: J
authors.lastname: Hemingway
authors.forename: Mark J I
authors.initials: MJ
authors.lastname: Paine
mesh_terms: D000818:Animals; D009033:Culicidae / Q000201:enzymology*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007305:Insecticide Resistance / Q000502:physiology; D007306:Insecticides / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D051379:Mice; D046228:Microarray Analysis; D015335:Molecular Probes / Q000378:metabolism*; D015394:Molecular Structure; D011722:Pyrethrins / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D013045:Species Specificity
chemical_list: D007306:Insecticides; D015335:Molecular Probes; D011722:Pyrethrins; D003577:Cytochrome P-450 Enzyme System
keywords: drug metabolism; insecticide resistance; interactome; malaria