Measurement of Phospholipid Metabolism in Intact Neutrophils

Phospholipid metabolizing enzymes are important participants in neutrophil signal transduction pathways. The methods discussed herein describe assays for assessing the activities of phospholipase (PL)A2 , PLC, PLD, and phosphoinositide 3-OH-kinase (PI3-K) in intact neutrophils. PLA2 activity is measured as the release of radiolabed arachidonic acid. PLC activity is measured as the accumulation of inositol 1,4,5-trisphosphate (IP3 ), a water-soluble product, using a commercially available radioreceptor assay kit. PLD activity is measured as the appearance of its radiolabeled products, phosphatidic acid and phosphatidylethanol. PI3-K activity is measured as the appearance of its radiolabeled product, phosphatidylinositol-3,4,5-trisphosphate.