Group V phospholipase A2 induces leukotriene biosynthesis in human neutrophils through the activation of group IVA phospholipase A2.
We reported previously that exogenously added human group V phospholipase A(2) (hVPLA(2)) could elicit leukotriene B(4) (LTB(4)) biosynthesis in human neutrophils (Han, S. K., Kim, K. P., Koduri, R., Bittova, L., Munoz, N. M., Leff, A. R., Wilton, D. C., Gelb, M. H., and Cho, W. (1999) J. Biol. Chem. 274, 11881-11888). To determine the mechanism of the hVPLA(2)-induced LTB(4) biosynthesis in neutrophils, we thoroughly examined the effects of hVPLA(2) and their lipid products on the activity of group IVA cytosolic PLA(2) (cPLA(2)) and LTB(4) biosynthesis under different conditions. As low as 1 nm exogenous hVPLA(2) was able to induce the release of arachidonic acid (AA) and LTB(4). Typically, AA and LTB(4) were released in two phases, which were synchronized with a rise in intracellular calcium concentration ([Ca(2+)](i)) near the perinuclear region and cPLA(2) phosphorylation. A cellular PLA(2) assay showed that hVPLA(2) acted primarily on the outer plasma membrane, liberating fatty acids and lysophosphatidylcholine (lyso-PC), whereas cPLA(2) acted on the perinuclear membrane. Lyso-PC and polyunsaturated fatty acids including AA activated cPLA(2) and 5-lipoxygenase by increasing [Ca(2+)](i) and inducing cPLA(2) phosphorylation, which then led to LTB(4) biosynthesis. The delayed phase was triggered by the binding of secreted LTB(4) to the cell surface LTB(4) receptor, which resulted in a rise in [Ca(2+)](i) and cPLA(2) phosphorylation through the activation of mitogen-activated protein kinase, extracellular signal-regulated kinase 1/2. These results indicate that a main role of exogenous hVPLA(2) in neutrophil activation and LTB(4) biosynthesis is to activate cPLA(2) and 5-lipoxygenase primarily by liberating from the outer plasma membrane lyso-PC that induces [Ca(2+)](i) increase and cPLA(2) phosphorylation and that hVPLA(2)-induced LTB(4) production is augmented by the positive feedback activation of cPLA(2) by LTB(4).
Department of Chemistry, University of Illinois at Chicago, Chicago, Illinois 60607, USA.
Young Jun
YJ
Kim
Kwang Pyo
KP
Kim
Sang Kyou
SK
Han
Nilda M
NM
Munoz
Xiangdong
X
Zhu
Hiroyuki
H
Sano
Alan R
AR
Leff
Wonhwa
W
Cho
D002118:Calcium / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D054513:Group IV Phospholipases A2; D054505:Group V Phospholipases A2; D006801:Humans; D006868:Hydrolysis; D015151:Immunoblotting; D015289:Leukotrienes / Q000096:biosynthesis*; D008244:Lysophosphatidylcholines / Q000378:metabolism; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000201:enzymology* / Q000378:metabolism; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011994:Recombinant Proteins / Q000378:metabolism; D013997:Time Factors
D015289:Leukotrienes; D008244:Lysophosphatidylcholines; D011994:Recombinant Proteins; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054505:Group V Phospholipases A2; C516389:PLA2G4A protein, human; D054467:Phospholipases A2; C101424:phospholipase A2-V, Trimeresurus; D002118:Calcium