Protein kinase A inhibits leukotriene synthesis by phosphorylation of 5-lipoxygenase on serine 523.
Leukotrienes (LTs) are lipid messengers generated by leukocytes that drive inflammation and modulate neighboring cell function. The synthesis of LTs from arachidonic acid is initiated by the enzyme 5-lipoxygenase (5-LO). We report for the first time that LT synthesis is inhibited by the direct action of protein kinase A (PKA) on 5-LO. The catalytic subunit of PKA directly phosphorylated 5-LO in vivo and in vitro and inhibited activity in intact cells and in vitro. Mutation of Ser-523 on human 5-LO prevented phosphorylation by PKA and restored LT synthesis. Treatment with PKA activators inhibited LTB(4) synthesis in 3T3 cells expressing wild type 5-LO but not in cells expressing the S523A mutant of 5-LO. The mechanism of inhibition of LTB(4) synthesis did not involve either reduced membrane association of activated 5-LO or redistribution of 5-LO from the nucleus to the cytoplasm. Instead, PKA phosphorylation of recombinant 5-LO inhibited in vitro activity, as did co-transfection of cells with 5-LO plus the catalytic subunit of PKA. Also, substitution of Ser-523 with glutamic acid, mimicking phosphorylation, resulted in the total loss of 5-LO activity. These results indicate that PKA phosphorylates 5-LO on Ser-523, which inhibits the catalytic activity of 5-LO and reduces cellular LT generation. Thus, PKA activation, as can occur in response to adenosine, prostaglandin E(2), beta-adrenergic agonists, and other mediators, is a means of directly reducing 5-LO activity and LT synthesis that may be important in limiting inflammation and maintaining homeostasis.
Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109, USA.
Ming
M
Luo
Sandra M
SM
Jones
Susan M
SM
Phare
Michael J
MJ
Coffey
Marc
M
Peters-Golden
Thomas G
TG
Brock
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D007975:Leukotriene B4 / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D015289:Leukotrienes / Q000096:biosynthesis*; D016859:Lipoxygenase Inhibitors; D051379:Mice; D020125:Mutation, Missense; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism; D014162:Transfection
D015289:Leukotrienes; D016859:Lipoxygenase Inhibitors; D007975:Leukotriene B4; D012694:Serine; D001094:Arachidonate 5-Lipoxygenase; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium