5-lipoxygenase is phosphorylated by p38 kinase-dependent MAPKAP kinases

O Werz, J Klemm, B Samuelsson… - Proceedings of the …, 2000 - National Acad Sciences
O Werz, J Klemm, B Samuelsson, O Rådmark
Proceedings of the National Academy of Sciences, 2000National Acad Sciences
5-lipoxygenase (5-LO) catalyzes the initial steps in the formation of leukotrienes, a group of
inflammatory mediators derived from arachidonic acid (AA). Here we describe that activation
of p38 mitogen-activated protein kinase in human polymorphonuclear leukocytes and in
Mono Mac 6 cells leads to activation of downstream kinases, which can subsequently
phosphorylate 5-LO in vitro. Different agents activated the 5-LO kinase activities, including
stimuli for cellular leukotriene biosynthesis (A23187, thapsigargin, N-formyl-leucyl …
5-lipoxygenase (5-LO) catalyzes the initial steps in the formation of leukotrienes, a group of inflammatory mediators derived from arachidonic acid (AA). Here we describe that activation of p38 mitogen-activated protein kinase in human polymorphonuclear leukocytes and in Mono Mac 6 cells leads to activation of downstream kinases, which can subsequently phosphorylate 5-LO in vitro. Different agents activated the 5-LO kinase activities, including stimuli for cellular leukotriene biosynthesis (A23187, thapsigargin, N-formyl-leucyl-phenylalanine), compounds that up-regulate the capacity for leukotriene biosynthesis (phorbol 12-myristate 13-acetate, tumor necrosis factor α, granulocyte/macrophage colony-stimulating factor), and well known p38 stimuli as sodium arsenite and sorbitol. For all stimuli, 5-LO kinase activation was counteracted by SB203580 (3 μM or less), an inhibitor of p38 kinase. At least two p38-dependent 5-LO kinase activities were found. Based on migration properties in in-gel kinase assays and immunoreactivity, one of these was identified as mitogen-activated protein kinase-activated protein kinase 2 (MAPKAP kinase 2). The other appeared to be MAPKAP kinase 3; however, it could not be excluded that also other p38-dependent kinases contributed. When polymorphonuclear leukocytes were incubated with sodium arsenite (strong activator of 5-LO kinases), platelet-activating factor and exogenous AA, there was a 4-fold increase in 5-LO activity as compared with incubations with only platelet-activating factor and AA. This indicates that 5-LO phosphorylation can be one factor determining cellular 5-LO activity.
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