Activation of human neutrophil nicotinamide adenine dinucleotide phosphate, reduced (triphosphopyridine nucleotide, reduced) oxidase by arachidonic acid in a cell-free system.

Curnutte, John T.

doi:10.1172/jci111885

Cited by 305 publications

(96 citation statements)

References 13 publications

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“…Many reports emphasize the effects of AA on p47 phox . Early studies showed AA or other anionic amphiphiles activate ROS production in cell-free assays of Nox2 activity (Bromberg and Pick, 1984;Curnutte, 1985). It is thought that AA induces conformational changes in p47 phox that may mimic the effects of phosphorylation, resulting in exposure of its binding PX and SH3 domains and translocation of p47 phox and p67 phox (Shiose and Sumimoto, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…However, recent studies in p40 phox -deficient mice or in Fc␥IIA receptor-reconstituted cells indicate p40 phox is an essential component of the Nox2 system (Ellson et al, 2006;Suh et al, 2006). Arachidonic acid (AA) has been used frequently as a activator of Nox2 in cell-free assay systems (Bromberg and Pick, 1984;Curnutte, 1985). Although relatively high concentrations of AA (50 -200 M) are required for activation of Nox2 both in vitro and in vivo, functional roles for cytosolic phospholipase A 2 (cPLA 2 ), which produces AA, in Nox2 activation have been demonstrated at the cellular level (Dana et al, 1998;Zhao et al, 2002;Shmelzer et al, 2003).…”

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confidence: 99%

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A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox

Ueyama

Tatsuno

Kawasaki

et al. 2007

MBoC

101

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In the phagocytic cell, NADPH oxidase (Nox2) system, cytoplasmic regulators (p47 phox , p67 phox , p40 phox , and Rac) translocate and associate with the membrane-spanning flavocytochrome b 558 , leading to activation of superoxide production. We examined membrane targeting of phox proteins and explored conformational changes in p40 phox that regulate its translocation to membranes upon stimulation. GFP-p40 phox translocates to early endosomes, whereas GFP-p47 phox translocates to the plasma membrane in response to arachidonic acid. In contrast, GFP-p67 phox does not translocate to membranes when expressed alone, but it is dependent on p40 phox and p47 phox for its translocation to early endosomes or the plasma membrane, respectively. Translocation of GFP-p40 phox or GFP-p47 phox to their respective membrane-targeting sites is abolished by mutations in their phox (PX) domains that disrupt their interactions with their cognate phospholipid ligands. Furthermore, GFP-p67 phox translocation to either membrane is abolished by mutations that disrupt its interaction with p40 phox or p47 phox . Finally, we detected a head-to-tail (PX-Phox and Bem1 [PB1] domain) intramolecular interaction within p40 phox in its resting state by deletion mutagenesis, cell localization, and binding experiments, suggesting that its PX domain is inaccessible to interact with phosphatidylinositol 3-phosphate without cell stimulation. Thus, both p40 phox and p47 phox function as diverse p67 phox "carrier proteins" regulated by the unmasking of membrane-targeting domains in distinct mechanisms. INTRODUCTIONIn phagocytic cells, reactive oxygen species (ROS) are produced by NADPH oxidase (Nox2 system). The enzyme is a multiprotein complex assembled from a membrane-spanning flavocytochrome b 558 (composed of gp91 phox [Nox2] and p22 phox ) and four cytoplasmic components (p47 phox , p67 phox , p40 phox , and Rac) (Leto, 1999;Nauseef, 2004;Quinn and Gauss, 2004). In unstimulated phagocytes, the oxidase is dissociated and inactive: the flavocytochrome b 558 is stored on the membranes of intracellular granules (Jesaitis et al., 1990), Rac is maintained in a GDP-bound cytoplasmic complex dimerized with Rho-guanine nucleotide dissociation inhibitor (GDI) , and the other phox proteins associate in a separate ternary cytoplasmic complex (p47 phox -p67 phox -p40 phox ) in a dephosphorylated state (Bolscher et al., 1989;Rotrosen and Leto, 1990;Kuribayashi et al., 2002;Lapouge et al., 2002). During phagocyte activation, intracellular granules containing flavocytochrome b 558 fuse with phagosomes; p47 phox is phosphorylated, thereby inducing conformational changes in p47 phox that promote the interaction of the cytoplasmic complex with the flavocytochrome b 558 ; and Rac dissociates from Rho-GDI and translocates independently to the membrane after exchange of GDP for GTP (Heyworth et al., 1994;Zhao et al., 2003), resulting in generation of superoxide anion by the transfer of electrons from cytoplasmic NADPH to molecular oxygen.Chronic granulomatous disease...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox

Ueyama

Tatsuno

Kawasaki

et al. 2007

MBoC

101

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“…These include the activation of protein kinases to phosphorylate cellular proteins and NADPH oxidase components (2,8) and the generation of various lipid second messengers (AA by phospholipase A 2 (9); DG by phospholipase C or PA phosphohydrolase; PA by phospholipase D or DG kinase (10,11)). In cell-free systems, these lipids can induce activation of the enzyme (12)(13)(14)(15)(16)(17). AA exerts its effect by directly acting on enzyme components (18 -22).…”

mentioning

confidence: 99%

Phosphatidic Acid and Diacylglycerol Directly Activate NADPH Oxidase by Interacting with Enzyme Components

Palicz

Foubert

Jesaitis

et al. 2001

Journal of Biological Chemistry

112

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The enzyme NADPH oxidase is regulated by phospholipase D in intact neutrophils and is activated by phosphatidic acid (PA) plus diacylglycerol (DG) in cell-free systems. We showed previously that cell-free NADPH oxidase activation by these lipids involves both protein kinase-dependent and -independent pathways. Here we demonstrate that only the protein kinase-independent pathway is operative in a cell-free system of purified and recombinant NADPH oxidase components. Activation by PA ؉ DG was ATP-independent and unaffected by the protein kinase inhibitor staurosporine, indicating the lack of protein kinase involvement. Both PA and DG were required for optimal activation to occur. The drug R59949 reduced activation of NADPH oxidase by either arachidonic acid or PA ؉ DG, with IC 50 values of 46 and 25 M, respectively. The optimal concentration of arachidonic acid or PA ؉ DG for oxidase activation was shifted to the right with R59949, indicating interference of the drug with the interaction of lipid activators and enzyme components. R59949 inhibited the lipid-induced aggregation/sedimentation of oxidase components p47 phox and p67 phox , suggesting a disruption of the lipidmediated assembly process. The direct effects of R59949 on NADPH oxidase activation complicate its use as a "specific" inhibitor of DG kinase. We conclude that the protein kinase-independent pathway of NADPH oxidase activation by PA and DG involves direct interaction with NADPH oxidase components. Thus, NADPH oxidase proteins are functional targets for these lipid messengers in the neutrophil.The NADPH oxidase (the respiratory burst enzyme) in phagocytic cells produces superoxide (O 2 . ) 1 by catalyzing electron transfer from NADPH to molecular oxygen upon cell stimulation (1-3). This enzyme plays important roles in host defense against infection and in tissue damage due to inflammation (1-5). In addition, NADPH oxidase-like enzymes are present in a variety of other cell types, where the oxygen radicals formed may have signaling roles (5, 6). The enzyme in phagocytes consists of the membrane-bound heterodimeric flavocytochrome b 558 (gp91 phox and p22 phox ) and four cytosolic proteins (p47 phox , p67 phox , p40 phox , Rac1/2) (2-4, 7). Components must assemble in the membrane for the enzyme to become active (2-4, 7). The activation of NADPH oxidase is initiated by receptor-ligand interaction and involves complex intracellular signaling events. These include the activation of protein kinases to phosphorylate cellular proteins and NADPH oxidase components (2, 8) and the generation of various lipid second messengers (AA by phospholipase A 2 (9); DG by phospholipase C or PA phosphohydrolase; PA by phospholipase D or DG kinase (10, 11)). In cell-free systems, these lipids can induce activation of the enzyme (12-17). AA exerts its effect by directly acting on enzyme components (18 -22). PA has been shown to partially activate purified flavocytochrome b 558 (23), suggesting it interacts with this protein. It is not known whether DG has any direct effe...

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“…27). Soon after that report, three other laboratories independently developed cell-free systems and confirmed that under very specific conditions, O 2̇̄g eneration could be supported by combining cytosol and membranes from resting phagocytes (28)(29)(30). Coupled with the ferricytochrome c assay, use of the cell-free system made possible the discovery of cytoplasmic proteins required for normal phagocyte oxidase activity (reviewed in Ref.…”

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confidence: 98%

Identification and Quantitation of Superoxide Anion: Essential Steps in Elucidation of the Phagocyte “Respiratory Burst”

Nauseef

2014

The Journal of Immunology

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Activation of human neutrophil nicotinamide adenine dinucleotide phosphate, reduced (triphosphopyridine nucleotide, reduced) oxidase by arachidonic acid in a cell-free system.

Cited by 305 publications

References 13 publications

A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox

A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox

Phosphatidic Acid and Diacylglycerol Directly Activate NADPH Oxidase by Interacting with Enzyme Components

Identification and Quantitation of Superoxide Anion: Essential Steps in Elucidation of the Phagocyte “Respiratory Burst”

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Activation of human neutrophil nicotinamide adenine dinucleotide phosphate, reduced (triphosphopyridine nucleotide, reduced) oxidase by arachidonic acid in a cell-free system.

Cited by 305 publications

References 13 publications

A Regulated Adaptor Function of p40phox: Distinct p67phoxMembrane Targeting by p40phoxand by p47phox

A Regulated Adaptor Function of p40phox: Distinct p67phoxMembrane Targeting by p40phoxand by p47phox

Phosphatidic Acid and Diacylglycerol Directly Activate NADPH Oxidase by Interacting with Enzyme Components

Identification and Quantitation of Superoxide Anion: Essential Steps in Elucidation of the Phagocyte “Respiratory Burst”

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A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox

A Regulated Adaptor Function of p40^phox: Distinct p67^phoxMembrane Targeting by p40^phoxand by p47^phox