1. The central role of mosquito cytochrome P450 CYP6Zs in insecticide detoxification revealed by functional expression and structural modelling
- Author
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Alexia Chandor-Proust, Jessica Roux, Muhammad Asam Riaz, Jean-Philippe David, Jaclyn Bibby, Mark J. I. Paine, Myriam Régent-Kloeckner, Chantal Dauphin-Villemant, Rodolphe Poupardin, Stéphane Reynaud, and Emilie Guittard-Crilat
- Subjects
cytochrome P450 monooxygenase ,0106 biological sciences ,Insecticides ,Anopheles gambiae ,recombinant system ,Benzoates ,01 natural sciences ,Biochemistry ,Insecticide Resistance ,chemistry.chemical_compound ,Cytochrome P-450 Enzyme System ,Aedes ,7-OH, 7-hydroxycoumarin ,Pyrethrins ,0303 health sciences ,Pyrethroid ,pyrethroid ,Cyt b5, cytochrome b5 ,Rt, retention time ,qPCR, quantitative real-time PCR ,Cytochrome P450 reductase ,PBA, 3-phenoxybenzoic acid ,3. Good health ,Genetically modified organism ,Isoenzymes ,Molecular Docking Simulation ,qw_52 ,qx_510 ,Benzaldehydes ,qu_26.5 ,Inactivation, Metabolic ,Insect Proteins ,Genetic Engineering ,Oxidation-Reduction ,Research Article ,In silico ,mosquito ,Saccharomyces cerevisiae ,Aedes aegypti ,Biology ,SRS, substrate recognition site ,Microbiology ,resistance ,TFA, trifluoroacetic acid ,03 medical and health sciences ,PBAld, 3-phenoxybenzaldehyde ,qx_600 ,Anopheles ,parasitic diseases ,Botany ,Escherichia coli ,Animals ,CPR, NADPH-cytochrome P450-reductase ,Aedes/drug effects ,Aedes/enzymology ,Anopheles gambiae/drug effects ,Anopheles gambiae/enzymology ,Benzaldehydes/chemistry ,Benzaldehydes/metabolism ,Benzoates/metabolism ,Cytochrome P-450 Enzyme System/chemistry ,Cytochrome P-450 Enzyme System/genetics ,Escherichia coli/enzymology ,Escherichia coli/genetics ,Insect Proteins/chemistry ,Insect Proteins/genetics ,Insect Vectors/drug effects ,Insect Vectors/enzymology ,Insecticide Resistance/drug effects ,Insecticide Resistance/genetics ,Insecticides/chemistry ,Insecticides/metabolism ,Isoenzymes/chemistry ,Isoenzymes/genetics ,Metabolic Detoxication, Drug ,NADPH-Ferrihemoprotein Reductase/chemistry ,NADPH-Ferrihemoprotein Reductase/genetics ,Pyrethrins/chemistry ,Pyrethrins/metabolism ,Saccharomyces cerevisiae/drug effects ,Saccharomyces cerevisiae/enzymology ,Molecular Biology ,NADPH-Ferrihemoprotein Reductase ,030304 developmental biology ,AeCPR, Aedes aegypti CPR ,PBAlc, 3-phenoxybenzoic alcohol ,insecticide ,wa_240 ,Cytochrome P450 ,Cell Biology ,Monooxygenase ,biology.organism_classification ,Insect Vectors ,010602 entomology ,chemistry ,biology.protein ,metabolism - Abstract
The resistance of mosquitoes to chemical insecticides is threatening vector control programmes worldwide. Cytochrome P450 monooxygenases (CYPs) are known to play a major role in insecticide resistance, allowing resistant insects to metabolize insecticides at a higher rate. Among them, members of the mosquito CYP6Z subfamily, like Aedes aegypti CYP6Z8 and its Anopheles gambiae orthologue CYP6Z2, have been frequently associated with pyrethroid resistance. However, their role in the pyrethroid degradation pathway remains unclear. In the present study, we created a genetically modified yeast strain overexpressing Ae. aegypti cytochrome P450 reductase and CYP6Z8, thereby producing the first mosquito P450–CPR (NADPH-cytochrome P450-reductase) complex in a yeast recombinant system. The results of the present study show that: (i) CYP6Z8 metabolizes PBAlc (3-phenoxybenzoic alcohol) and PBAld (3-phenoxybenzaldehyde), common pyrethroid metabolites produced by carboxylesterases, producing PBA (3-phenoxybenzoic acid); (ii) CYP6Z8 transcription is induced by PBAlc, PBAld and PBA; (iii) An. gambiae CYP6Z2 metabolizes PBAlc and PBAld in the same way; (iv) PBA is the major metabolite produced in vivo and is excreted without further modification; and (v) in silico modelling of substrate–enzyme interactions supports a similar role of other mosquito CYP6Zs in pyrethroid degradation. By playing a pivotal role in the degradation of pyrethroid insecticides, mosquito CYP6Zs thus represent good targets for mosquito-resistance management strategies., The key role of mosquito cytochrome P450 monooxygenases from the CYP6Z subfamily in the resistance of mosquitoes to insecticides was revealed. By using functional expression and in silico modelling, the capacity of CYP6Zs to degrade insecticide metabolites produced by esterase-mediated hydrolysis was demonstrated.
- Published
- 2013