Premium
The role of diacylglyceride generation by phospholipase D and phosphatidic acid phosphatase in the activation of 5‐lipoxygenase in polymorphonuclear leukocytes
Author(s) -
Albert Dana,
Pergola Carlo,
Koeberle Andreas,
Dodt Gabriele,
Steinhilber Dieter,
Werz Oliver
Publication year - 2008
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1189/jlb.0807531
Subject(s) - phosphatidic acid , lipoxygenase , biology , phospholipase d , acid phosphatase , phospholipase , biochemistry , phosphatase , phospholipase c , enzyme , microbiology and biotechnology , phospholipid , membrane
Diacylglycerides (DAGs) such as 1‐oleoyl‐2‐acetyl‐sn‐glycerol (OAG) stimulate 5‐lipoxygenase (5‐LO) enzyme activity and function as agonists for human polymorphonuclear leukocytes (PMNL) to induce 5‐LO product synthesis. Here, we addressed the role of endogenous DAG generation in agonist‐induced 5‐LO activation in human PMNL. Preincubation of PMNL with the phospholipase D (PLD) inhibitor 1‐butanol potently suppressed 5‐LO product synthesis induced by the Ca 2 + ionophore A23187 or thapsigargin (TG) and blocked A23187‐evoked translocation of 5‐LO from the cytosol to the nuclear membrane, analyzed by subcellular fractionation as well as by indirect immunofluorescence microscopy. Tertiary‐butanol, a rather poor inhibitor of PLD, caused only moderate suppression of 5‐LO and hardly inhibited 5‐LO translocation. Interestingly, 1‐butanol failed to inhibit 5‐LO product formation when PMNL were stimulated with OAG (30 μM). Moreover, coincubation of A23187‐ or TG‐stimulated PMNL with OAG reversed inhibition of 5‐LO product formation by 1‐butanol in a concentration‐dependent manner (EC 50 , ∼1 μM) and also restored 5‐LO translocation. In addition, inhibition of phosphatidic acid phosphatase (PA‐P) by propranolol or bromoenol lactone caused suppression of 5‐LO product formation and of translocation, which could be reversed by addition of exogenous OAG. Together, our data suggest that in agonist‐stimulated PMNL, the endogenous formation of DAGs via the PLD/PA‐P pathway determines 5‐LO activation.