Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids

Jiawen Xu, Christophe Morisseau, Bruce D. Hammock

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In insects, epoxide hydrolases (EHs) play critical roles in the metabolism of xenobiotic epoxides from the food resources and in the regulation of endogenous chemical mediators, such as juvenile hormones. Using the baculovirus expression system, we expressed and characterized an epoxide hydrolase from Anopheles gambiae (AgEH) that is distinct in evolutionary history from insect juvenile hormone epoxide hydrolases (JHEHs). We partially purified the enzyme by ion exchange chromatography and isoelectric focusing. The experimentally determined molecular weight and pI were estimated to be 35kD and 6.3 respectively, different than the theoretical ones. The AgEH had the greatest activity on long chain epoxy fatty acids such as 14,15-epoxyeicosatrienoic acids (14,15-EET) and 9,10-epoxy-12Z-octadecenoic acids (9,10-EpOME or leukotoxin) among the substrates evaluated. Juvenile hormone III, a terpenoid insect growth regulator, was the next best substrate tested. The AgEH showed kinetics comparable to the mammalian soluble epoxide hydrolases, and the activity could be inhibited by AUDA [12-(3-adamantan-1-yl-ureido) dodecanoic acid], a urea-based inhibitor designed to inhibit the mammalian soluble epoxide hydrolases. The rabbit serum generated against the soluble epoxide hydrolase of Mus musculus can both cross-react with natural and denatured forms of the AgEH, suggesting immunologically they are similar. The study suggests there are mammalian sEH homologs in insects, and epoxy fatty acids may be important chemical mediators in insects.

Original languageEnglish (US)
Pages (from-to)42-52
Number of pages11
JournalInsect Biochemistry and Molecular Biology
Volume54
DOIs
StatePublished - Nov 1 2014

Fingerprint

epoxide hydrolase
Anopheles gambiae
Epoxide Hydrolases
epoxides
Fatty Acids
fatty acids
juvenile hormones
Insects
Juvenile Hormones
insects
Insect Hormones
leukotoxins
insect hormones
Baculoviridae
Epoxy Compounds
Terpenes
Ion Exchange Chromatography
Isoelectric Focusing
Xenobiotics
Substrates

Keywords

  • Anopheles gambiae
  • Characterization
  • Epoxide hydrolase
  • Epoxy fatty acids

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

Cite this

Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids. / Xu, Jiawen; Morisseau, Christophe; Hammock, Bruce D.

In: Insect Biochemistry and Molecular Biology, Vol. 54, 01.11.2014, p. 42-52.

Research output: Contribution to journalArticle

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