A novel assay system implicates PtdIns(3,4)P(2), PtdIns(3)P, and PKC delta in intracellular production of reactive oxygen species by the NADPH oxidase.

Academic Article

Abstract

  • Activated neutrophils assemble an NADPH oxidase enzyme complex to produce superoxide for microbial killing. Much of the initial oxidase assembly occurs on intracellular granules, followed by movement of the oxidase to phagolysosomes and the plasma membrane. We have developed a novel assay system using Streptolysin-O permeabilized neutrophils that recapitulates the initial intracellular activation process while maintaining the ultrastructural features of this granulocytic cell type. Using this system, we biochemically dissect molecular events and signaling pathways involved in NADPH oxidase assembly and demonstrate specific roles for PKC delta, PI(3,4)P(2)/PI(3,4,5)P(3), and PI(3)P in the PMA-dependent intracellular activation process. This system should be of great utility for the study of cell signaling events that regulate the intracellular production of reactive oxygen species by neutrophils.
  • Authors

  • Brown, Glenn E
  • Stewart, Mary
  • Liu, Hui
  • Ha, Vi-Luan
  • Yaffe, Michael B
  • Status

    Publication Date

  • January 2003
  • Published In

  • Molecular Cell  Journal
  • Keywords

  • Adenosine Triphosphate
  • Bacterial Proteins
  • Cell Fractionation
  • Cytoplasm
  • Enzyme Inhibitors
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Humans
  • NADPH Oxidases
  • Neutrophil Activation
  • Neutrophils
  • Phosphatidylinositol 3-Kinases
  • Phosphatidylinositol Phosphates
  • Phosphoproteins
  • Protein Kinase C
  • Protein Kinase C-delta
  • Protein Transport
  • Reactive Oxygen Species
  • Signal Transduction
  • Streptolysins
  • Tetradecanoylphorbol Acetate
  • Digital Object Identifier (doi)

    Pubmed Id

  • 12535519
  • Start Page

  • 35
  • End Page

  • 47
  • Volume

  • 11
  • Issue

  • 1