Activation of p38 in Stimulated Human Neutrophils: Phosphorylation of the Oxidase Component p47 by p38 and ERK but Not by JNK

Benna, Jamel El; Han, Jiahuai; Park, Jeen‐Woo; Schmid, Elmar; Ulevitch, Richard J.; Babior, Bernard M.

doi:10.1006/abbi.1996.0470

Cited by 231 publications

(178 citation statements)

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“…In addition, the p38-kinase inhibitor SB-203580 totally inhibited phosphorylation and translocation of p47 phox and superoxide production. The demonstration that, in vitro, p38 kinase can phosphorylate p47 phox strengthens our hypothesis (36). Our results also suggest that p38-kinase activation in As 2 O 3 -treated macrophages involved the serine-threonine kinase ROCK; indeed, the ROCK inhibitor Y-27632 was found to prevent phosphorylation of p38 kinase and to mimic inhibitory effects of SB-203580 on NADPH oxidase activation: it markedly reduced phosphorylation of p47 phox , its membrane translocation, and superoxide production.…”

Section: Discussionsupporting

confidence: 87%

Inorganic Arsenic Activates Reduced NADPH Oxidase in Human Primary Macrophages through a Rho Kinase/p38 Kinase Pathway

Lemarié

Bourdonnay

Morzadec

et al. 2008

The Journal of Immunology

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Inorganic arsenic is an immunotoxic environmental contaminant to which millions of humans are chronically exposed. We recently demonstrated that human primary macrophages constituted a critical target for arsenic trioxide (As2O3), an inorganic trivalent form. To specify the effects of arsenic on macrophage phenotype, we investigated in the present study whether As2O3 could regulate the activity of NADPH oxidase, a major superoxide-generating enzymatic system in human phagocytes. Our results show that superoxide levels were significantly increased in a time-dependent manner in blood monocyte-derived macrophages treated with 1 μM As2O3 for 72 h. Concomitantly, As2O3 induced phosphorylation and membrane translocation of the NADPH oxidase subunit p47phox and it also increased translocation of Rac1 and p67phox. Apocynin, a selective inhibitor of NADPH oxidases, prevented both p47phox translocation and superoxide production. NADPH oxidase activation was preceded by phosphorylation of p38-kinase in As2O3-treated macrophages. The p38-kinase inhibitor SB-203580 prevented phosphorylation and translocation of p47phox and subsequent superoxide production. Pretreatment of macrophages with the Rho-kinase inhibitor Y-27632 was found to mimic inhibitory effects of SB-203580 and to prevent As2O3-induced phosphorylation of p38 kinase. Treatment with As2O3 also resulted in an increased secretion of the proinflammatory chemokine CCL18 that was fully inhibited by both apocynin and SB-203580. Taken together, our results demonstrate that As2O3 induced a marked activation of NADPH oxidase in human macrophages, likely through stimulation of a Rho-kinase/p38-kinase pathway, and which may contribute to some of the deleterious effects of inorganic arsenic on macrophage phenotype.

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Section: Discussionsupporting

confidence: 87%

Inorganic Arsenic Activates Reduced NADPH Oxidase in Human Primary Macrophages through a Rho Kinase/p38 Kinase Pathway

Lemarié

Bourdonnay

Morzadec

et al. 2008

The Journal of Immunology

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“…It has been described that ERK and p38 MAPK phosphorylate p47 phox in human neutrophils [41] and that inhibitors of these kinases attenuate the neutrophil respiratory burst stimulated by a wide array of agonists [42,43]. Moreover, our group has shown that angiotensin II-dependent O ÅÀ 2 production becomes suppressed by specific inhibitors of the p38MAPK and ERK1/2 pathways [29].…”

Section: Resultsmentioning

confidence: 86%

Stimulators of AMP‐activated protein kinase inhibit the respiratory burst in human neutrophils

et al. 2004

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In the present study, we have examined the potential ability of 5 0 -AMP-activated protein kinase (AMPK) to modulate NADPH oxidase activity in human neutrophils. AMPK activated with either 5 0 -aminoimidazole-4-carboxamide ribonucleoside (AICAR) or with 5 0 -AMP significantly attenuated both phorbol 12-myristate 13-acetate (PMA) and formyl methionyl leucyl phenylalanine-stimulated superoxide anion ðO À 2 Þ release by human neutrophils, consistently with a reduced translocation to the cell membrane and phosphorylation of a cytosolic component of NADPH oxidase, namely p47 phox . AMPK was found to be present in human neutrophils and to become phosphorylated in response to either AICAR or other stimulators of its enzyme activity. Furthermore, AICAR also strongly reduced PMA-dependent H 2 O 2 release, and induced the phosphorylation of c-jun N-terminal kinase 1 (p46), p38 mitogenactivated protein kinase and extracellular signal-regulated kinase. Present data demonstrate for the first time that the activation of AMPK, in states of low cellular energy charge (such as under high levels of 5 0 -AMP) or other signals, could be a factor contributing to reduce the host defense mechanisms.

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“…Understanding the molecular mechanisms by which p38 MAPK participates in these responses is hindered by the limited number of targets of p38 MAPK identified to date in neutrophils. MAP-KAPK2 and p47 phox are the only clearly identified p38 MAPK targets in human neutrophils (1,9,10,21). A previous study by Lewis et al (45) determined that 20 of 25 ERK targets identified in a global screen were not previously known.…”

Section: Discussionmentioning

confidence: 97%

“…A number of p38 MAPK substrates have been identified previously in other cell types, including transcription factors such as activating transcription factor 2, kinases such as p38-regulated and -activated kinase and MAPKactivated protein kinase 2 (MAPKAPK-2), 3 and cell cycle regulators such as cyclin D (17)(18)(19)(20). The only p38 MAPK substrates that have been identified in human neutrophils, however, are MAP-KAPK-2 and p47 phox (1,9,10,21). The goal of the present study was to identify p38 MAPK substrates in human neutrophils using a functional proteomic approach developed in our laboratory that combines in vitro phosphorylation of neutrophil lysate by recombinant kinase, separation of proteins by two-dimensional gel electrophoresis, and phosphoprotein identification by MALDI-TOF mass spectrometry (MALDI-TOF MS) (22,23).…”

mentioning

confidence: 99%

Myeloid-Related Protein-14 Is a p38 MAPK Substrate in Human Neutrophils

Lominadze

Rane

Merchant

et al. 2005

The Journal of Immunology

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The targets of the p38 MAPK pathway that mediate neutrophil functional responses are largely unknown. To identify p38 MAPK targets, a proteomic approach was applied in which recombinant active p38 MAPK and [32P]ATP were added to lysates from unstimulated human neutrophils. Proteins were separated by two-dimensional gel electrophoresis, and phosphoproteins were visualized by autoradiography and identified by MALDI-TOF. Myeloid-related protein-14 (MRP-14) was identified as a candidate p38 MAPK substrate. MRP-14 phosphorylation by p38 MAPK was confirmed by an in vitro kinase reaction using purified MRP-14/MRP-8 complexes. The site of MRP-14 phosphorylation by p38 MAPK was identified by tandem mass spectrometry and site-directed mutagenesis to be Thr113. MRP-14 phosphorylation by p38 MAPK in intact neutrophils was confirmed by [32P]orthophosphate loading, followed by fMLP stimulation in the presence and absence of a p38 MAPK inhibitor, SB203580. Confocal microscopy of Triton X-100 permeabilized neutrophils showed that a small amount of MRP-14 was associated with cortical F-actin in unstimulated cells. fMLP stimulation resulted in a p38 MAPK-dependent increase in MRP-14 staining at the base of lamellipodia. By immunoblot analysis, MRP-14 was present in plasma membrane/secretory vesicle fractions and gelatinase and specific granules, but not in azurophil granules. The amount of MRP-14 associated with plasma membrane/secretory vesicle and gelatinase granule fractions increased after fMLP stimulation in a p38 MAPK-dependent manner. Direct phosphorylation of the MRP-14/MRP-8 complex by p38 MAPK increased actin binding in vitro by 2-fold. These results indicate that MRP-14 is a potential mediator of p38 MAPK-dependent functional responses in human neutrophils.

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Activation of p38 in Stimulated Human Neutrophils: Phosphorylation of the Oxidase Component p47 by p38 and ERK but Not by JNK

Cited by 231 publications

References 0 publications

Inorganic Arsenic Activates Reduced NADPH Oxidase in Human Primary Macrophages through a Rho Kinase/p38 Kinase Pathway

Inorganic Arsenic Activates Reduced NADPH Oxidase in Human Primary Macrophages through a Rho Kinase/p38 Kinase Pathway

Stimulators of AMP‐activated protein kinase inhibit the respiratory burst in human neutrophils

Myeloid-Related Protein-14 Is a p38 MAPK Substrate in Human Neutrophils

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