Impaired p47phox phosphorylation in neutrophils from p67phox-deficient chronic granulomatous disease patients Short title: p67phox regulates p47phox phosphorylation - Archive ouverte HAL
Article Dans Une Revue Blood Année : 2022

Impaired p47phox phosphorylation in neutrophils from p67phox-deficient chronic granulomatous disease patients Short title: p67phox regulates p47phox phosphorylation

Shiyu Liang
  • Fonction : Auteur
Yezhou Liu
Richard Ye

Résumé

Abstract Superoxide production by the phagocyte reduced NAD phosphate (NADPH) oxidase is essential for innate immunity as shown in chronic granulomatous disease (CGD), an immunodeficiency disease resulting from mutations in 1 of its genes. The NADPH oxidase is composed of 2 membrane proteins (gp91phox/NOX2 and p22phox) and 4 cytosolic proteins (p47phox, p67phox, p40phox, and Rac1/2). The phosphorylation of p47phox is required for NADPH oxidase activation in cells. As p47phox and p67phox can form a tight complex in cells, we hypothesized that p67phox could regulate p47phox phosphorylation. To investigate this hypothesis, we used phospho-specific antibodies against 5 major p47phox-phosphorylated sites (Ser304, Ser315, Ser320, Ser328, and Ser345) and neutrophils from healthy donors and from p67phox−/− CGD patients. Results showed that formyl-methionyl-leucyl-phenylalanine and phorbol myristate acetate induced a time- and a concentration-dependent phosphorylation of p47phox on Ser304, Ser315, Ser320, and Ser328 in healthy human neutrophils. Interestingly, in neutrophils and Epstein-Barr virus-transformed B lymphocytes from p67phox−/− CGD patients, phosphorylation of p47phox on serine residues was dramatically reduced. In COSphox cells, the presence of p67phox led to increased phosphorylation of p47phox. In vitro studies showed that recombinant p47phox was phosphorylated on Ser304, Ser315, Ser320, and Ser328 by different PKC isoforms and the addition of recombinant p67phox alone or in combination with p40phox potentiated this process. Thus, p67phox and p40phox are required for optimal p47phox phosphorylation on Ser304, Ser315, Ser320, and Ser328 in intact cells. Therefore, p67phox and p40phox are novel regulators of p47phox-phosphorylation.

Domaines

Immunité innée
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Dates et versions

hal-03810711 , version 1 (11-10-2022)

Identifiants

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Sahra Amel Belambri, Viviana Marzaioli, Margarita Hurtado-Nedelec, Coralie Pintard, Shiyu Liang, et al.. Impaired p47phox phosphorylation in neutrophils from p67phox-deficient chronic granulomatous disease patients Short title: p67phox regulates p47phox phosphorylation. Blood, 2022, 139 (16), pp.2512-2522. ⟨10.1182/blood.2021011134⟩. ⟨hal-03810711⟩
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