Involvement of protein kinase D in Fcγ-receptor activation of the NADPH oxidase in neutrophils
Involvement of protein kinase D in Fcγ-receptor activation of the NADPH oxidase in neutrophils
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DOI：
10.1042/0264-6021:3630095
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发表时间：
2002-04-01
影响因子：
4.1
通讯作者：
Dekker, LV
中科院分区：
文献类型：
--
作者：
Davidson-Moncada, JK;Lopez-Lluch, G;Dekker, LV
研究了中性粒细胞中参与Fcgamma受体激活NADPH氧化酶的蛋白激酶。在三种不同的PKC抑制剂中，Go 6976完全抑制NADPH氧化酶，而双吲哚酰亚胺I和Ro 31-8220的抑制作用为70-80%。因此，一个对Go 6976敏感、双吲哚酰马来酰亚胺I/Ro 31-8220不敏感的组分有助于Fcgamma受体诱导的NADPH氧化酶激活。PKC同型的下调导致fcgamma受体激活的NADPH氧化酶的抑制，但仍然存在下调不敏感的成分。该组件对Go 6976敏感，但对Ro 31-8220不敏感。先前已有研究表明，蛋白激酶D/PKC-mu （PKD）在体外表现出相同的药理作用。我们发现PKD存在于中性粒细胞中。与PKC同型相比，PKD没有下调。因此PKD可能参与NADPH氧化酶的活化。为了获得直接证据，我们采用了反义方法。反义PKD对fcγ受体刺激诱导的NADPH氧化酶的抑制作用为50%，活化过程中Ro 31-8220不敏感成分被反义PKD抑制。体外激酶实验表明，PKD通过将igg活化的颗粒呈献给中性粒细胞而被激活。此外，PKD定位于细胞中颗粒摄入的区域，并磷酸化NADPH氧化酶的三种细胞质成分中的两种，p40（phox）和p47（phox）。综上所述，这些数据表明Fcgamma受体参与PKD对NADPH氧化酶的调节。
Protein kinases involved in the activation of the NADPH oxidase by Fcgamma receptors in neutrophils were studied. Of three different protein kinase C (PKC) inhibitors, Go 6976 inhibited the NADPH oxidase completely, whereas bisindolylmateimide I and Ro 31-8220 caused a 70-80% inhibition. Thus a Go 6976-sensitive, bisindolylmaleimide I/Ro 31-8220-insensitive component contributes to NADPH oxidase activation induced by Fcgamma receptors. Down-regulation of PKC isotypes resulted in inhibition of Fcgamma-receptor-activated NADPH oxidase, but a down-regulation-insensitive component was still present. This component was sensitive to Go 6976, but insensitive to Ro 31-8220. It has been shown previously that protein kinase D/PKC-mu (PKD) shows this same pharmacology in vitro. We show that PKD is present in neutrophils and that., in contrast with PKC isotypes, PKD is not down-regulated. Therefore PKD may participate in NADPH oxidase activation. To obtain direct evidence for this we adopted an antisense approach. Antisense PKD inhibited NADPH oxidase induced by Fcgamma-receptor stimulation by 50% and the Ro 31-8220-insensitive component in the activation was inhibited by antisense PKD. In vitro kinase assays showed that PKD is activated by presenting IgG-opsonized particles to neutrophils, Furthermore, PKD localizes to the area of particle intake in the cell and phosphorylates two of the three cytosolic components of the NADPH oxidase, p40(phox) and p47(phox). Taken together, these data indicate that Fcgamma receptors engage PKD in the regulation of the NADPH oxidase.