Lipid A stimulates phospholipase D activity in rat mesangial cells via a G-protein.

Abstract

Stimulation of mesangial cells (MC) with the bacterial endotoxin Lipid A activated two enzymes involved in lipid metabolism. First, a phospholipase D hydrolyses phosphatidylethanolamine (PE) to phosphatidic acid (PA), followed by dephosphorylation of PA to 1,2-diacylglycerol (DAG) by PA phosphohydrolase. MC or microsomes from these cells were pre-labelled with [3H]glycerol. A 30-60 s stimulation with 10-100 ng of Lipid A/ml caused a decrease in [3H]glycerol in PE and increased radioactive glycerol in PA. The enzyme responsible for this hydrolysis preferred PE containing unsaturated acyl side chains. DAG was formed from PA within the first 1 min after Lipid A stimulation. Microsomes incubated with 25 mM-NaF to inhibit phospholipase C and to stimulate GTP-binding proteins also caused PE to be converted into PA. The [3H]glycerol and acyl mass of phosphatidylcholine, phosphatidylserine and phosphatidylinositol did not change with either Lipid A or NaF. Addition of guanosine 5'-[gamma-thio]triphosphate to MC microsomes caused the rapid decrease in proportion of PE and increase in PA, followed by an increase in DAG unsaturated acyl mass. These data suggest the concurrent G-protein-dependent activation by Lipid A of a PE-directed phospholipase D and a PA phosphohydrolase.