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: E M Van der Donk, G R Dubois, J Verhagen, G A Veldink, J F Vliegenthart
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1074 Issue 3 Pg. 443-7 (Aug 06 1991) ISSN: 0006-3002 [Print] Netherlands
• PMID: 1888756 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Lipoxygenase Inhibitors
• Arachidonate 12-Lipoxygenase
• Arachidonate 5-Lipoxygenase
• Calcium

Topics

• Animals
• Arachidonate 12-Lipoxygenase (isolation & purification, metabolism)
• Arachidonate 5-Lipoxygenase (metabolism)
• Calcium (metabolism)
• Chromatography, High Pressure Liquid
• Hydrogen-Ion Concentration
• Leukemia, Basophilic, Acute (enzymology)
• Lipoxygenase Inhibitors
• Rats
• Tumor Cells, Cultured (enzymology)