title: Biosynthesis of eoxin C4 by porcine leukocytes.
abstract: Human 15-lipoxygenase-1 (LO) possesses mainly 15-lipoxygenase activity whereas the animal ortholog 12/15-LO possesses mainly 12-lipoxygenase activity. These findings have raised the question if studies on animals can predict the function of 15-LO-1 in human. In this study we have characterized the arachidonic acid metabolites formed by porcine 12/15-LO. Mini pigs were infected with a parasite to increase the number of blood eosinophils, which highly express 12/15-LO. Isolated porcine polymorphonuclear leukocytes (PMNL) were incubated with arachidonic acid and the produced metabolites were analysed with HPLC and mass spectrometry (MS). The cells were found to produce 15-hydroxyeicosatetraenoic acid (HETE) and 12-HETE at a ratio of 1:5. Furthermore 8,15-dihydroxyeicosatetraenoic acids (DiHETEs) and 14,15-DiHETE were formed. Based on HPLC, UV-spectroscopy and MS analysis it was found that porcine PMNL also produced eoxin (EX) C4. These results demonstrate that although porcine 12/15-LO possesses primarily 12-lipoxygenase activity, the enzyme can catalyse the formation of EXC(4).
authors.affiliation: Department of Medicine, Karolinska University Hospital and Karolinska Institutet, Solna, SE-171 76 Stockholm, Sweden. asa.brunnstrom@ki.se
authors.forename: Åsa
authors.initials: Å
authors.lastname: Brunnström
authors.forename: Linda
authors.initials: L
authors.lastname: Backman
authors.forename: Ylva
authors.initials: Y
authors.lastname: Tryselius
authors.forename: Hans-Erik
authors.initials: HE
authors.lastname: Claesson
mesh_terms: D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000378:metabolism*; D016718:Arachidonic Acid / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D007962:Leukocytes / Q000378:metabolism*; D015289:Leukotrienes / Q000096:biosynthesis*; D013058:Mass Spectrometry; D009504:Neutrophils / Q000378:metabolism; D013552:Swine
chemical_list: C475064:12-15-lipoxygenase; D015289:Leukotrienes; C526311:eoxin C4; D016718:Arachidonic Acid; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase