Phosphorylation- and stimulus-dependent inhibition of cellular 5-lipoxygenase activity by nonredox-type inhibitors

FASEB J. 2003 May;17(8):949-51. doi: 10.1096/fj.02-0815fje. Epub 2003 Mar 28.


Nonredox-type 5-lipoxygenase (5-LO) inhibitors such as ZM230487 or L-739.010 potently suppress leukotriene biosynthesis at low cellular peroxide tone. Here, we show that inhibition of 5-LO product formation by nonredox-type 5-LO inhibitors in human isolated polymorphonuclear leukocytes (PMNL) depends on the activation pathway of 5-LO. Thus, compared with 5-LO product synthesis induced by the Ca2+-mobilizing agent ionophore A23187, cell stress-induced 5-LO product formation involving 5-LO kinase pathways required ~10- to 100-fold higher concentrations of ZM230487 or L-739.010 for comparable 5-LO inhibition. No such differences were observed for the iron ligand-type 5-LO inhibitor BWA4C or the novel-type 5-LO inhibitors hyperforin and 3-O-acetyl-11-keto-boswellic acid. Experiments using purified 5-LO revealed that Ca2+ is no prerequisite for potent enzyme inhibition by ZM230487, and exposure of PMNL to the combination of ionophore and cell stress did not restore potent 5-LO suppression. Intriguingly, a significant difference in the potency of nonredox-type inhibitors (but not of BWA4C) was determined between wild-type 5-LO and the mutant S271A/S663A-5-LO (lacking phosphorylation sites for ERK1/2 and MAPKAPK-2) in HeLa cells. Collectively, our data suggest that compared with Ca2+-mediated 5-LO product formation, enzyme activation involving 5-LO phosphorylation events specifically and strongly alters the susceptibility of 5-LO toward nonredox-type inhibitors in intact cells.

MeSH terms

  • Arachidonate 5-Lipoxygenase / genetics
  • Arachidonate 5-Lipoxygenase / metabolism
  • Arsenites / pharmacology
  • Benzeneacetamides*
  • Bridged Bicyclo Compounds / pharmacology
  • Calcimycin / pharmacology
  • Calcium / metabolism
  • Dose-Response Relationship, Drug
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology*
  • HeLa Cells
  • Humans
  • Hydroxamic Acids / pharmacology
  • Lipoxygenase Inhibitors* / pharmacology
  • Mitogen-Activated Protein Kinases / metabolism
  • Mutation, Missense
  • Neutrophils / cytology
  • Neutrophils / drug effects*
  • Neutrophils / metabolism
  • Peroxides / metabolism
  • Phloroglucinol / analogs & derivatives
  • Phosphorylation
  • Pyrans / pharmacology
  • Quinolines / pharmacology
  • Quinolones / pharmacology
  • Sodium Chloride / pharmacology
  • Sodium Compounds / pharmacology
  • Terpenes / pharmacology


  • Arsenites
  • Benzeneacetamides
  • Bridged Bicyclo Compounds
  • Enzyme Inhibitors
  • Hydroxamic Acids
  • L 739010
  • Lipoxygenase Inhibitors
  • Peroxides
  • Pyrans
  • Quinolines
  • Quinolones
  • Sodium Compounds
  • Terpenes
  • N-(3-phenoxycinnamyl)acetohydroxamic acid
  • ZM 230487
  • Calcimycin
  • Sodium Chloride
  • sodium arsenite
  • Phloroglucinol
  • Arachidonate 5-Lipoxygenase
  • Mitogen-Activated Protein Kinases
  • hyperforin
  • Calcium