title: Improved purification of 12-lipoxygenase from rat basophilic leukemia cells and conditions for optimal enzyme activity.
abstract: 12-Lipoxygenase from rat basophilic leukemia cells was purified about 300-fold by protein-HPLC in a single run. Maximal 12-lipoxygenase activity was observed at pH 7.5, while the enzyme became almost inactive at pH 6 and 9. Although Ca2+ was not essential for 12-lipoxygenase activity, the partially purified enzyme was stimulated approx. 2-fold in the presence of 0.1-5.0 mM Ca2+. Contrary to 5-lipoxygenase from RBL-1 cells, 12-lipoxygenase was not inactivated by preincubation with Ca2+ for 1-10 min, nor was it stimulated by 0.1-10 mM ATP.
authors.affiliation: Bijvoet Center for Biomolecular Research, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
authors.forename: E M
authors.initials: EM
authors.lastname: Van der Donk
authors.forename: G R
authors.initials: GR
authors.lastname: Dubois
authors.forename: J
authors.initials: J
authors.lastname: Verhagen
authors.forename: G A
authors.initials: GA
authors.lastname: Veldink
authors.forename: J F
authors.initials: JF
authors.lastname: Vliegenthart
mesh_terms: D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000302:isolation & purification* / Q000378:metabolism; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D015471:Leukemia, Basophilic, Acute / Q000201:enzymology*; D016859:Lipoxygenase Inhibitors; D051381:Rats; D014407:Tumor Cells, Cultured / Q000201:enzymology
chemical_list: D016859:Lipoxygenase Inhibitors; D001092:Arachidonate 12-Lipoxygenase; D001094:Arachidonate 5-Lipoxygenase; D002118:Calcium