Regulation of 5-lipoxygenase activity in mononuclear phagocytes: characterization of an endogenous cytosolic inhibitor.
The proinflammatory leukotrienes (LT) play important roles in host defense and disease states. However, no endogenous mechanisms to downregulate 5-lipoxygenase (5-LO), the enzyme catalyzing LT synthesis, have been described. We observed that the cytosolic fraction of rat alveolar macrophages (AMs) and peritoneal macrophages (PMs), and of peripheral blood monocytes (PBMs) contain substantial amounts of 5-LO protein, but little detectable 5-LO activity. We therefore examined these mononuclear phagocyte (MNP) cytosolic fractions for inhibitory activity against 5-LO. MNP cytosol dose-dependently reduced the 5-LO activity in neutrophil (PMN) cytosol and AM membrane. Furthermore, MNP cytosol dose-dependently prolonged the lag phase of soybean lipoxygenase (LO) without affecting the rate of product formation. This effect was overcome by subsequent addition of 13(S)-hydroperoxy-9-cis-11-trans-octadecadienoic acid (13-HpOD), suggesting that the active factor scavenges hydroperoxides. Inactivation by boiling and roteinase K suggest that is a protein. We speculate that this cytosolic factor(s) may serve as an endogenous means for the down-regulation of 5-LO in macrophages.
Divisions of Pulmonary and Critical Care Medicine, University of Michigan Medical Center, Ann Arbor 48109, USA. mcoffey@umich.edu
M J
MJ
Coffey
S E
SE
Wilcoxen
P H
PH
Sporn
M
M
Peters-Golden
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D002478:Cells, Cultured; D003600:Cytosol / Q000737:chemistry* / Q000201:enzymology; D006706:Homeostasis; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015151:Immunoblotting; D016859:Lipoxygenase Inhibitors / Q000032:analysis*; D016676:Macrophages, Alveolar / Q000201:enzymology / Q000648:ultrastructure; D017737:Macrophages, Peritoneal / Q000201:enzymology / Q000648:ultrastructure; D009000:Monocytes / Q000201:enzymology / Q000648:ultrastructure; D009504:Neutrophils / Q000201:enzymology; D010586:Phagocytes / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D013025:Glycine max / Q000201:enzymology
D016859:Lipoxygenase Inhibitors; D006861:Hydrogen Peroxide; D001094:Arachidonate 5-Lipoxygenase