Linoleic acid peroxidation by Solanum tuberosum lipoxygenase was activated in the presence of human 5-lipoxygenase-activating protein.
The present investigation describes the ability of human 5-lipoxygenase-activating protein (FLAP) to activate a plant 5-lipoxygenase. The presence of an active recombinant human FLAP in the 100000xg membrane fraction of infected Sf9 cells led to a specific increase in 9-hydroperoxyoctadecadienoic acid (9-HPOD) synthesis (+68%) or in 5-hydroperoxyeicosatetraenoic acid (5-HPETE) synthesis (+68%), after action of Solanum tuberosum tuber 5-lipoxygenase (S.t.LOX) on linoleic acid (natural plant lipoxygenase substrate) or on arachidonic acid. On the contrary, the presence of non-transfected membranes obtained from non-infected Sf9 cells led to an inhibition of lipoxygenase activity. MK-886, a potent inhibitor of leukotriene biosynthesis, blocked the FLAP dependent S.t.LOX activation after preincubation with FLAP transfected membranes. In conclusion, this study demonstrates that a recombinant human FLAP can stimulate a lipoxygenase other than mammalian 5-lipoxygenase (S.t.LOX) by using different polyunsaturated fatty acids as substrates.
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Laboratoires de Biochimie, Facultés de Médecine et de Pharmacie, 2 rue du Docteur Marcland, 87025 Limoges Cedex, France.
Moalic
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Rigaud
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Beneytout
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JL
D058927:5-Lipoxygenase-Activating Proteins; D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D016718:Arachidonic Acid; D016367:Baculoviridae / Q000235:genetics; D002118:Calcium / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics / Q000494:pharmacology*; D002462:Cell Membrane / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D015289:Leukotrienes / Q000096:biosynthesis; D019787:Linoleic Acid / Q000378:metabolism*; D008041:Linoleic Acids / Q000096:biosynthesis; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000494:pharmacology; D011198:Solanum tuberosum / Q000201:enzymology*; D018411:Spodoptera / Q000378:metabolism / Q000648:ultrastructure; D014162:Transfection; D002722:Chlorobenzenes
D058927:5-Lipoxygenase-Activating Proteins; D001094:Arachidonate 5-Lipoxygenase; D016718:Arachidonic Acid; D002118:Calcium; D002352:Carrier Proteins; D007211:Indoles; D015289:Leukotrienes; D019787:Linoleic Acid; D008041:Linoleic Acids; D016859:Lipoxygenase Inhibitors; D008565:Membrane Proteins; D011994:Recombinant Proteins; C548592:ALOX5AP protein, human; C060893:MK-886; D002722:Chlorobenzenes; C049606:9-hydroperoxy-11,12-octadecadienoic acid; C025348:arachidonic acid 5-hydroperoxide