Structural organization of the neutrophil NADPH oxidase: phosphorylation and translocation during priming and activation

  title={Structural organization of the neutrophil NADPH oxidase: phosphorylation and translocation during priming and activation},
  author={Forest R. Sheppard and Marguerite R. Kelher and Ernest E. Moore and Nathan J. D. McLaughlin and Anirban Banerjee and Christopher C Silliman},
  journal={Journal of Leukocyte Biology},
The reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is part of the microbicidal arsenal used by human polymorphonuclear neutrophils (PMNs) to eradicate invading pathogens. The production of a superoxide anion (O2–) into the phagolysosome is the precursor for the generation of more potent products, such as hydrogen peroxide and hypochlorite. However, this production of O2– is dependent on translocation of the oxidase subunits, including gp91phox, p22phox, p47phox, p67phox… 

Assessment of priming of the human neutrophil respiratory burst.

The techniques used to measure priming of the NADPH oxidase in human neutrophils are described, which are a "double-edged sword" process as it contributes to a rapid and efficient elimination of the pathogens but can also induce the generation of large quantities of toxic ROS that can damage surrounding tissues and participate to inflammation.

The Prolyl Isomerase Pin1 Controls Lipopolysaccharide-Induced Priming of NADPH Oxidase in Human Neutrophils

Results suggest that the prolyl cis/trans isomerase Pin1 may control LPS-induced priming of superoxide production in human neutrophils and Pharmacological targeting of Pin1 could be a valuable approach in sepsis.

PtdIns3P and Rac direct the assembly of the NADPH oxidase on a novel, pre-phagosomal compartment during FcR-mediated phagocytosis in primary mouse neutrophils.

A novel pathway for oxidase assembly downstream of FcR-activation is defined,defined, by using a variety of assays to follow the spatiotemporal assembly of the oxidase in genetically engineered primary mouse neutrophils, during phagocytosis of both serum- and immunoglobulin G-opsonized targets.

NADPH oxidase activity: In the crossroad of neutrophil life and death.

A growing number of evidence suggests that NAD PHox-derived ROS are involved in the activation of signaling pathways that may lead to increased neutrophil survival, highlighting the signaling pathways modulated by NADPHox- derived ROS.

Neutrophils from p40phox−/− mice exhibit severe defects in NADPH oxidase regulation and oxidant-dependent bacterial killing

The defect in ROS production by p40phox−/− neutrophils in response to S. aureus translated into a severe, CGD-like defect in the killing of this organism both in vitro and in vivo, defining p40 phox as an essential component in bacterial killing.

Modulating the activity of NADPH oxidase by oxidative stress participants ; lipids and nanoparticles A cell-free system study

This work studied the NADPH oxidase functioning in an in vitro system in which the components of the enzyme are mixed and activated by the introduction of an amphiphile the arachidonic acid (AA), and showed that the cholesterol already present in the phagocyte membrane is optimal for the function NADPH oxidation.

Priming of the neutrophil NADPH oxidase activation: role of p47phox phosphorylation and NOX2 mobilization to the plasma membrane

Some of the mechanisms of NADPH oxidase priming in neutrophils and the relevance of this process to physiology and pathology are discussed.

trans Arachidonic acid isomers inhibit NADPH-oxidase activity by direct interaction with enzyme components.

Evaluation of p47phox phosphorylation in human neutrophils using phospho-specific antibodies.

A radioactive-free technique to analyze the phosphorylation of p47phox in cell lysates, based on the use of phospho-specific antibodies, SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and Western blotting is described.



Activation of NADPH oxidase of human neutrophils involves the phosphorylation and the translocation of cytosolic p67phox.

The immunoprecipitated p67phox from neutrophil cytosol and membrane fractions demonstrate that the phosphorylation of p67 phox is correlated with activation of NADPH oxidase, and continuous phosphorylated of p 67phox is required in order to maintain the linearity of the respiratory burst.

Cytosolic phox proteins interact with and regulate the assembly of coronin in neutrophils.

The cytosolic phox proteins might play a regulatory role in the reorganisation of the cytoskeleton accompanying superoxide generation and the rearrangement of F-actin and coronin in adherent cells.

Regulation of phagocyte oxygen radical production by the GTP-binding protein Rac 2.

The results suggest that Rac 2 is a regulatory component of the human neutrophil NADPH oxidase, and provide new insights into the mechanism by which this oxygen radical-generating system is regulated.

The superoxide-generating oxidase of phagocytic cells. Physiological, molecular and pathological aspects.

The present review is focused on recent data concerning the signaling pathway which leads to oxidase activation, including specific receptors, the production of second messengers, the organization of the oxidase complex and the molecular defects responsible for granulomatous disease.

The 40-kDa component of the phagocyte NADPH oxidase (p40phox) is phosphorylated during activation in differentiated HL60 cells.

In this study, it is shown using dimethylsulfoxide-differentiated HL60 promyelocytes that p40phox is in a basal phosphorylated state in resting cells and undergoes further phosphorylation on multiple sites upon stimulation of the NADPH oxidase by either phorbol myristate acetate or by the formyl peptide fMet-Leu-Phe-Lys.

Mechanisms of NADPH oxidase activation in human neutrophils: p67phox is required for the translocation of rac 1 but not of rac 2 from cytosol to the membranes.

This is the first demonstration that, in activated neutrophils, rac 1 is translocated from the cytosol to the membranes and this translocation requires p67phox, and may suggest that the assembly of the cytOSolic components of NADPH oxidase on the plasma membrane takes place through selective coupling of activated rac 1 and rac 2 with p67 phox and p47phox respectively.

Assembly of the phagocyte NADPH oxidase: molecular interaction of oxidase proteins

The key features of each NADPH oxidase protein are described and the current understanding of the specific molecular interactions occurring between these proteins are summarized, focusing on the role these protein:protein binding interactions play in the NADPH oxidation assembly process.