title: A novel protein kinase target for the lipid second messenger phosphatidic acid.
abstract: Activation of phospholipase D occurs in response to a wide variety of hormones, growth factors, and other extracellular signals. The initial product of phospholipase D, phosphatidic acid (PA), is thought to serve a signaling function, but the intracellular targets for this lipid second messenger are not clearly identified. The production of PA in human neutrophils is closely correlated with the activation of NADPH oxidase, the enzyme responsible for the respiratory burst. We have developed a cell-free system, in which the activation of NADPH oxidase is induced by the addition of PA. Characterization of this system revealed that a multi-functional cytosolic protein kinase was a target for PA, and that two NADPH oxidase components were substrates for the enzyme. Partial purification of the PA-activated protein kinase separated the enzyme from known protein kinase targets of PA. The partially purified enzyme was selectively activated by PA, compared to other phospholipids, and phosphorylated the oxidase component p47-phox on both serine and tyrosine residues. PA-activated protein kinase activity was present in a variety of hematopoietic cells and cell lines and in rat brain, suggesting it has widespread distribution. We conclude that this protein kinase may be a novel target for the second messenger function of PA.
authors.affiliation: Department of Biochemistry, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC, 27157-1019, USA. lmcphail@wfubmc.edu
authors.forename: L C
authors.initials: LC
authors.lastname: McPhail
authors.forename: K A
authors.initials: KA
authors.lastname: Waite
authors.forename: D S
authors.initials: DS
authors.lastname: Regier
authors.forename: J B
authors.initials: JB
authors.lastname: Nixon
authors.forename: D
authors.initials: D
authors.lastname: Qualliotine-Mann
authors.forename: W X
authors.initials: WX
authors.lastname: Zhang
authors.forename: R
authors.initials: R
authors.lastname: Wallin
authors.forename: S
authors.initials: S
authors.lastname: Sergeant
mesh_terms: D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D026901:Membrane Transport Proteins; D009252:NADPH Dehydrogenase / Q000378:metabolism; D019255:NADPH Oxidases / Q000096:biosynthesis; D009504:Neutrophils / Q000378:metabolism; D010712:Phosphatidic Acids / Q000096:biosynthesis / Q000494:pharmacology / Q000502:physiology*; D010739:Phospholipase D / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D016897:Respiratory Burst; D015290:Second Messenger Systems
chemical_list: D026901:Membrane Transport Proteins; D010712:Phosphatidic Acids; D010750:Phosphoproteins; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase; D011494:Protein Kinases; D010739:Phospholipase D