The mitogen-activated protein kinase extracellular signal-regulated kinase 1/2 pathway is involved in formyl-methionyl-leucyl-phenylalanine-induced p47phox phosphorylation in human neutrophils
The mitogen-activated protein kinase extracellular signal-regulated kinase 1/2 pathway is involved in formyl-methionyl-leucyl-phenylalanine-induced p47phox phosphorylation in human neutrophils
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DOI：
10.4049/jimmunol.165.9.5238
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发表时间：
2000-11-01
影响因子：
4.4
通讯作者：
El-Benna, J
中科院分区：
文献类型：
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作者：
Dewas, C;Fay, M;El-Benna, J
p47吞噬细胞氧化酶（p47phox）是NADPH氧化酶的组分之一，其磷酸化对于该酶的激活以及超氧化物的产生至关重要。p47phox在多个丝氨酸残基上被磷酸化，但体内参与这一过程的激酶仍有待鉴定。我们研究了细胞外信号调节激酶（ERK1/2）和p38丝裂原活化蛋白激酶在p47phox磷酸化中的作用。用ERK激酶1/2的特异性抑制剂PD98059抑制ERK1/2的激活，可抑制甲酰甲硫氨酰亮氨酰苯丙氨酸（fMLP）诱导的p47phox磷酸化。然而，尽管ERK1/2的激活被阻断，但PD98059对佛波醇酯（PMA）诱导的p47phox磷酸化影响较弱。使用另一种ERK激酶抑制剂U0126也证实了这一效应。与PD98059和U0126不同，p38丝裂原活化蛋白激酶抑制剂SB203580不抑制fMLP或PMA诱导的p47phox磷酸化。二维磷酸肽图谱分析表明，在fMLP诱导的p47phox磷酸化中，PD98059影响所有主要磷酸肽的磷酸化，这表明ERK1/2可能直接或通过其他激酶间接调节p47phox磷酸化。在PMA诱导的p47phox磷酸化中，蛋白激酶C抑制剂GF109203X强烈抑制p47phox磷酸化。然而，在fMLP诱导的p47phox磷酸化中，PD98059和GF109203X单独使用时部分抑制p47phox磷酸化，一起使用时对p47phox磷酸化产生累加抑制作用。这些结果首次表明ERK1/2通路参与p47phox的磷酸化。此外，它们有力地表明，在由fMLP激活的完整中性粒细胞中，p47phox是几个激酶级联的作用靶点，因此是ERK1/2和蛋白激酶C的汇聚点。
Phosphorylation of p47 phagocyte oxidase, (p47(phox)), one of the NADPH oxidase components, is essential for the activation of this enzyme and for superoxide production. p47(phox) is phosphorylated on multiple serine residues, but the kinases involved in this process in vivo remain to be Characterized. We examined the role of extracellular signal-regulated kinase (ERK1/2) and p38 mitogen-activated protein kinase in p47(phox) phosphorylation, Inhibition of ERK1/2 activation by PD98059, a specific inhibitor of ERK kinase 1/2, inhibited the fMLP-induced phosphorylation of p47(phox). However, PD98059 weakly affected PMA-induced p47(phox) phosphorylation, even though ERK1/2 activation was abrogated. This effect was confirmed using U0126, a second ERK kinase inhibitor, Unlike PD98059 and U0126, the p38 mitogen-activated protein kinase inhibitor SB203580 did not inhibit the phosphorylation of p47(phox) induced either by fMLP or by PMA, Two-dimensional phosphopeptide mapping analysis showed that, in fMLP-induced p47(phox) phosphorylation, PD98059 affected the phosphorylation of all the major phosphopeptides, suggesting that ERK1/2 may regulate p47(phox) phosphorylation either directly or indirectly via other kinases, In PMA-induced p47(phox) phosphorylation, GF109203X a protein kinase C inhibitor, strongly inhibits p47(phox) phosphorylation. However, in fMLP-induced p47(phox) phosphorylation, PD98059 and GF109203X partially inhibited the phosphorylation of p47(phox) when tested alone, and exerted additive inhibitory effects on p47(phox) phosphorylation when tested together. These results show for the first time that the ERK1/2 pathway participates in the phosphorylation of p47(phox). Furthermore, they strongly suggest that p47(phox) is targeted by several kinase cascades in intact neutrophils activated by fMLP and is therefore a converging point for ERK1/2 and protein kinase C.