Affiliation:
1. Inositide Laboratory, Babraham Institute, Cambridge, United Kingdom;
2. Cardiovascular Division, Kings College London, London, United Kingdom; and
3. UCB Celltech, Cambridge, United Kingdom
Abstract
AbstractThe neutrophil nicotinamide adenine dinucleotide phosphate-oxidase is a multisubunit enzyme (comprising gp91phox, p22phox, p67phox, p40phox, p47phox, and Rac) that plays a vital role in microbial killing. The recent discovery of a chronic granulomatous disease patient who expresses a mutant p40phox subunit, together with the development of mouse models of p40phox function, indicate phosphatidylinositol 3-phosphate binding to the PX domain of p40phox is an important signal for oxidase activation. However, the presence of other conserved residues and domains in p40phox suggest further regulatory roles for this protein. To test this, we introduced wild-type and mutated versions of p40phox into fully differentiated mouse neutrophils by retroviral transduction of p40phox−/− bone marrow progenitors and repopulation of the bone marrow compartment in radiation chimaeras. Phosphorylation of p40phox on threonine 154, but not serine 315, was required for full oxidase activation in response to formylated bacterial peptide fMLP, serum-opsonized S aureus, and immunoglobulin-opsonized sheep red blood cells. A functional SH3 domain was not required for oxidase activation, and deletion of the entire domain resulted in enhanced oxidase responses. Phosphorylation of threonine 154 in response to S aureus was mediated by protein kinase Cδ and was required for full translocation of p47phox to phagosomes. These results define an important new element in the physiological activation of the oxidase.
Publisher
American Society of Hematology
Subject
Cell Biology,Hematology,Immunology,Biochemistry
Reference51 articles.
1. Neutrophils and immunity: challenges and opportunities.;Nathan;Nat Rev Immunol,2006
2. Inside the neutrophil phagosome: oxidants, myeloperoxidase, and bacterial killing.;Hampton;Blood,1998
3. Activation of NADPH oxidase involves the dissociation of p21rac from its inhibitory GDP/GTP exchange protein (rhoGDI) followed by its translocation to the plasma membrane.;Abo;Biochem J,1994
4. Phosphorylation of Rho GDI stabilizes the Rho A-Rho GDI complex in neutrophil cytosol.;Bourmeyster;Biochem Biophys Res Commun,1996
5. 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.;Brown;Mol Cell,2003
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