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 (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.
authors.affiliation: Department of Biochemistry, Wake Forest University, School of Medicine, Winston-Salem, NC, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Sergeant
authors.forename: Linda C
authors.initials: LC
authors.lastname: McPhail
mesh_terms: D000818:Animals; D002855:Chromatography, Thin Layer / Q000379:methods; D019411:Clinical Laboratory Techniques; D006801:Humans; D008954:Models, Biological; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000032:analysis / Q000378:metabolism; D010739:Phospholipase D / Q000032:analysis / Q000378:metabolism; D054467:Phospholipases A2 / Q000032:analysis / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis* / Q000378:metabolism*; D010738:Type C Phospholipases / Q000032:analysis / Q000378:metabolism
chemical_list: D010743:Phospholipids; D019869:Phosphatidylinositol 3-Kinases; D054467:Phospholipases A2; D010738:Type C Phospholipases; D010739:Phospholipase D