title: [Identification of lipoxygenase expressing omega-6 dioxygenase activity by means of liquid chromatography].
abstract: Lipoxygenases (LOX) represent a family of lipid peroxidising enzymes which catalyse the reaction of achiral polyunsaturated fatty acids by oxygen forming chiral peroxide products possessing high positional stereospecific purity. The four double bonds of arachidonic acid, the main substrate of animal LOX, present the position for a wide range of enzymatic modifications enabling eicosanoid creation, unique molecules with biological significance. In this study, lipoxygenase from rat lung cytoplasma was isolated and purified to 40-fold by combining hydrophobic and gel chromatography. The forming positional specific fatty acid hydroxyl-isomers were separated on a nonpolar system (RP-HPLC) and identified on a polar adsorbent (SP-HPLC). In the purified enzyme, dual positional specificity was demonstrated by the production of 12- and 15-HETE in the ratio of 1,0:1,38, which responds to the product spectrum of mammalian 15-LOX-1.
authors.affiliation: Katedra bunkovej a molekulárnej biológie lieciv, Farmaceutická fakulta Univerzita Komenského, Kalinciakova 8, 832 32 Bratislava, Slovenská republika. hoffman@fpharm.uniba.sk
authors.forename: Peter
authors.initials: P
authors.lastname: Hoffman
authors.forename: Mária
authors.initials: M
authors.lastname: Cupáková
authors.forename: Drahomíra
authors.initials: D
authors.lastname: Rauová
authors.forename: Renáta
authors.initials: R
authors.lastname: Kollárová
authors.forename: Mária
authors.initials: M
authors.lastname: Pekárová
authors.forename: Marek
authors.initials: M
authors.lastname: Oblozinský
authors.forename: Lýdia
authors.initials: L
authors.lastname: Bezáková
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D049308:Dioxygenases / Q000737:chemistry*; D008084:Lipoxygenase / Q000737:chemistry* / Q000302:isolation & purification; D008168:Lung / Q000201:enzymology; D051381:Rats
chemical_list: D049308:Dioxygenases; C572466:omega-6 dioxygenase, rat; D008084:Lipoxygenase