Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids期刊界 All Journals 搜尽天下杂志传播学术成果专业期刊搜索期刊信息化学术搜索

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

Institution:	3. Biosciences National Laboratory, National Center for Research in Energy and Materials, Rua Giuseppe Maximo Scolfaro 10000, 13083-100 Campinas/SP, Brazil;4. Proteomics and Protein Engineering Laboratory, Carlos Chagas Institute, Fiocruz, Rua Professor Algacyr Munhoz Mader 2135, 81310-020 Curitiba/PR, Brazil;5. i3S-Institute for Investigation and Innovation in Health, University of Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal;6. Institute of Molecular and Cell Biology (IBMC), University of Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal;12. Abel Salazar Biomedical Sciences Institute, University of Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal

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 35 kD 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.

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