title: Cloning and characterization of a microsomal epoxide hydrolase from Heliothis virescens.
abstract: Epoxide hydrolases (EHs) are α/β-hydrolase fold superfamily enzymes that convert epoxides to 1,2-trans diols. In insects EHs play critical roles in the metabolism of toxic compounds and allelochemicals found in the diet and for the regulation of endogenous juvenile hormones (JHs). In this study we obtained a full-length cDNA, hvmeh1, from the generalist feeder Heliothis virescens that encoded a highly active EH, Hv-mEH1. Of the 10 different EH substrates that were tested, Hv-mEH1 showed the highest specific activity (1180 nmol min(-1) mg(-1)) for a 1,2-disubstituted epoxide-containing fluorescent substrate. This specific activity was more than 25- and 3900-fold higher than that for the general EH substrates cis-stilbene oxide and trans-stilbene oxide, respectively. Although phylogenetic analysis placed Hv-mEH1 in a clade with some lepidopteran JH metabolizing EHs (JHEHs), JH III was a relatively poor substrate for Hv-mEH1. Hv-mEH1 showed a unique substrate selectivity profile for the substrates tested in comparison to those of MsJHEH, a well-characterized JHEH from Manduca sexta, and hmEH, a human microsomal EH. Hv-mEH1 also showed unique enzyme inhibition profiles to JH-like urea, JH-like secondary amide, JH-like primary amide, and non-JH-like primary amide compounds in comparison to MsJHEH and hmEH. Although Hv-mEH1 is capable of metabolizing JH III, our findings suggest that this enzymatic activity does not play a significant role in the metabolism of JH in the caterpillar. The ability of Hv-mEH1 to rapidly hydrolyze 1,2-disubstituted epoxides suggests that it may play roles in the metabolism of fatty acid epoxides such as those that are commonly found in the diet of Heliothis.
authors.affiliation: Department of Entomology and UC Davis Comprehensive Cancer Center, University of California, Davis, CA 95616, USA.
authors.forename: Shizuo G
authors.initials: SG
authors.lastname: Kamita
authors.forename: Kohji
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Mary M
authors.initials: MM
authors.lastname: Dadala
authors.forename: Khavong
authors.initials: K
authors.lastname: Pha
authors.forename: Christophe
authors.initials: C
authors.lastname: Morisseau
authors.forename: Aurélie
authors.initials: A
authors.lastname: Escaich
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Hammock
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004851:Epoxide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism*; D007605:Juvenile Hormones / Q000378:metabolism; D007814:Larva / Q000201:enzymology; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology*; D011869:Radioligand Assay; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D019476:Insect Proteins; D007605:Juvenile Hormones; D011994:Recombinant Proteins; D004851:Epoxide Hydrolases