The phosphorylation of the respiratory burst oxidase component p47phox during neutrophil activation. Phosphorylation of sites recognized by protein kinase C and by proline-directed kinases.
The phosphorylation of the respiratory burst oxidase component p47phox during neutrophil activation. Phosphorylation of sites recognized by protein kinase C and by proline-directed kinases.
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发表时间：
1994-09
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通讯作者：
J. Benna;L. R. Faust;B. Babior
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作者：
J. Benna;L. R. Faust;B. Babior
呼吸爆发氧化酶催化氧气和NADPH产生O2.-。它在休眠的细胞中处于休眠状态，但在细胞受到刺激时变得活跃。伴随着激活的是胞浆氧化酶组分p47Phox中的多个丝氨酸的磷酸化，在氧化酶激活过程中，它从胞浆移动到细胞膜。用佛波醇肉豆蔻酸醋酸酯激活的32PI负载的中性粒细胞的免疫纯化的p47Phox表明，所有的32P都在蛋白质的C端CNBR片段中，在该片段中，Ser-303、Ser-304、Ser-320、Ser-328、Ser-345和Ser-348以及三种丝氨酸中的至少一种Ser-359、Ser-370和Ser-379被磷酸化，而Ser-282、Ser-287、Ser-381和Ser-388没有被磷酸化。在磷酸化丝氨酸中，Ser-303、Ser-304、Ser-320和Ser-328位于蛋白激酶C底物序列中。然而，Ser-345和Ser-348位于丝裂原活化蛋白(MAP)激酶(-PXSP-)识别的序列中。这一发现表明，MAP激酶或相关的脯氨酸导向的激酶可能参与了激活的中性粒细胞产生O2.-的调节。经N-甲酰-甲硫基-亮氨酰苯丙氨酸活化的中性粒细胞p47Phox磷酸肽的胰酶肽谱与佛波醇活化细胞的p47Phox磷酸肽非常相似，表明相同的丝氨酸在不同的试剂作用下被磷酸化。
The respiratory burst oxidase catalyzes the production of O2.- from oxygen and NADPH. It is dormant in resting cells but becomes active when the cells are stimulated. Activation is accompanied by the phosphorylation of multiple serines in the cytosolic oxidase component p47phox, which moves from cytosol to the membrane during oxidase activation. Using immunopurified p47phox isolated from 32Pi-loaded neutrophils activated with phorbol myristate acetate, we showed that all the 32P was in the C-terminal CNBr fragment of the protein, and that in that fragment, Ser-303, Ser-304, Ser-320, Ser-328, Ser-345, and Ser-348 and at least one of the three serines, Ser-359, Ser-370, and Ser-379, were phosphorylated, while Ser-282, Ser-287, Ser-381, and Ser-388 were not. Of the phosphorylated serines, Ser-303, Ser-304, Ser-320, and Ser-328 are located in protein kinase C substrate sequences. Ser-345 and Ser-348, however, are located in sequences recognized by mitogen-activated protein (MAP) kinase (-PXSP-). This finding suggests that MAP kinase or a related proline-directed kinase may participate in the regulation of O2.- production by activated neutrophils. The tryptic peptide map of p47phox phosphopeptides from neutrophils activated by N-formyl-methionyl-leucyl-phenylalanine closely resembled that of p47phox phosphopeptides from phorbol-activated cells, suggesting that the same serines were phosphorylated in response to each agent.