Remarkable substrate-specificity of CYP6AB3 in Depressaria pastinacella, a highly specialized caterpillar

W. Mao, S. Rupasinghe, A. R. Zangerl, Mary A Schuler, May R Berenbaum

Research output: Contribution to journalArticle

Abstract

The parsnip webworm, Depressaria pastinacella, a specialist on two genera in Apiaceae, feeds exclusively on the furanocoumarin-containing reproductive structures of its host plants. This caterpillar relies principally on cytochrome P450-mediated detoxification for coping with the high concentrations of furanocoumarins in its diet. A cDNA encoding the furanocoumarin-inducible P450 CYP6AB3 from this species was coexpressed with house-fly NADPH P450 reductase in baculovirus-infected Sf9 cells and tested for binding and metabolism of the six furanocoumarins typically encountered in host plant tissues. Only imperatorin and bergapten bind in close proximity to the catalytic haem and only imperatorin is metabolized (Vmax and Km of 2.412 pmol/min per pmol P450 and 94.28 μm, respectively). Purification of the imperatorin metabolite by normal phase HPLC and characterization of its structure by MS-MS analysis indicate that CYP6AB3 initially epoxidizes the carbon-carbon pi-bond on the isoprenyl side chain on imperatorin. An improved molecular model for the CYP6AB3 protein based on this biochemical characterization and the recently defined mammalian CYP3A4 crystal structure provides insight into the remarkable substrate specificity of this protein.

Original languageEnglish (US)
Pages (from-to)169-179
Number of pages11
JournalInsect Molecular Biology
Volume15
Issue number2
DOIs
StatePublished - Apr 1 2006

Fingerprint

Depressaria pastinacella
psoralens
substrate specificity
Substrate Specificity
insect larvae
Pastinaca
Carbon
host plants
bergapten
Apiaceae
Sf9 Cells
Cytochrome P-450 CYP3A
NADPH-Ferrihemoprotein Reductase
molecular models
Molecular Models
carbon
Baculoviridae
Musca domestica
crystal structure
Heme

Keywords

  • Cytochrome P450 monooxygenases
  • Furanocoumarin metabolism
  • Homology modelling
  • Imperatorin
  • Plant-insect interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Insect Science

Cite this

Remarkable substrate-specificity of CYP6AB3 in Depressaria pastinacella, a highly specialized caterpillar. / Mao, W.; Rupasinghe, S.; Zangerl, A. R.; Schuler, Mary A; Berenbaum, May R.

In: Insect Molecular Biology, Vol. 15, No. 2, 01.04.2006, p. 169-179.

Research output: Contribution to journalArticle

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