title: Measurement of phospholipid metabolism in intact neutrophils.
abstract: Phospholipid-metabolizing enzymes are important participants in neutrophil signal transduction pathways. The methods discussed herein describe assays for assessing the activities of phospholipase A2 (PLA2), phospholipase C (PLC), phospholipase D (PLD), and phosphoinositide 3-OH-kinase in intact neutrophils. PLA2 activity is measured as the release of radiolabeled 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.
authors: Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC, USA.
authors: Susan
authors: S
authors: Sergeant
authors: Linda C
authors: LC
authors: McPhail
mesh_terms: D002855:Chromatography, Thin Layer / Q000379:methods; D004789:Enzyme Activation; D054513:Group IV Phospholipases A2 / Q000378:metabolism; D006801:Humans; D050356:Lipid Metabolism; D009504:Neutrophils / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D015398:Signal Transduction; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D010743:Phospholipids; D019869:Phosphatidylinositol 3-Kinases; D054513:Group IV Phospholipases A2; D010738:Type C Phospholipases; D010739:Phospholipase D