Heterologous expression and biochemical characterisation of fourteen esterases from Helicoverpa armigera

Mark G. Teese, Claire A. Farnsworth, Yongqiang Li, Chris W. Coppin, Alan L. Devonshire, Colin Scott, Peter East, Robyn J. Russell, John G. Oakeshott

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Abstract

Esterases have recurrently been implicated in insecticide resistance in Helicoverpa armigera but little is known about the underlying molecular mechanisms. We used a baculovirus system to express 14 of 30 full-length esterase genes so far identified from midgut cDNA libraries of this species. All 14 produced esterase isozymes after native PAGE and the isozymes for seven of them migrated to two regions of the gel previously associated with both organophosphate and pyrethroid resistance in various strains. Thirteen of the enzymes obtained in sufficient yield for further analysis all showed tight binding to organophosphates and low but measurable organophosphate hydrolase activity. However there was no clear difference in activity between the isozymes from regions associated with resistance and those from elsewhere in the zymogram, or between eight of the isozymes from a phylogenetic clade previously associated with resistance in proteomic and quantitative rtPCR experiments and five others not so associated. By contrast, the enzymes differed markedly in their activities against nine pyrethroid isomers and the enzymes with highest activity for the most insecticidal isomers were from regions of the gel and, in some cases, the phylogeny that had previously been associated with pyrethroid resistance. Phospholipase treatment confirmed predictions from sequence analysis that three of the isozymes were GPI anchored. This unusual feature among carboxylesterases has previously been suggested to underpin an association that some authors have noted between esterases and resistance to the Cry1Ac toxin from Bacillus thuringiensis. However these three isozymes did not migrate to the zymogram region previously associated with Cry1Ac resistance.

Original languageEnglish
Article numbere65951
Number of pages9
JournalPLoS ONE
Volume8
Issue number6
DOIs
Publication statusPublished - Jun 2013
Externally publishedYes

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Copyright the Author(s) 2013. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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    Teese, M. G., Farnsworth, C. A., Li, Y., Coppin, C. W., Devonshire, A. L., Scott, C., ... Oakeshott, J. G. (2013). Heterologous expression and biochemical characterisation of fourteen esterases from Helicoverpa armigera. PLoS ONE, 8(6), [e65951]. https://doi.org/10.1371/journal.pone.0065951