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首页> 外文期刊>Molecular cell >A Novel Assay System Implicates PtdIns(3,4)P(2), PtdIns(3)P, and PKCdelta in Intracellular Production of Reactive Oxygen Species by the NADPH Oxidase.
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A Novel Assay System Implicates PtdIns(3,4)P(2), PtdIns(3)P, and PKCdelta in Intracellular Production of Reactive Oxygen Species by the NADPH Oxidase.

机译:一种新颖的测定系统涉及NADPH氧化酶在胞内生产活性氧中的PtdIns(3,4)P(2),PtdIns(3)P和PKCdelta。

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摘要

Activated neutrophils assemble an NADPH oxidase enzyme complex to produce superoxide for microbial killing. Much of the initial oxidase assembly occurs on intracellular granules, followed by movement of the oxidase to phagolysosomes and the plasma membrane. We have developed a novel assay system using Streptolysin-O permeabilized neutrophils that recapitulates the initial intracellular activation process while maintaining the ultrastructural features of this granulocytic cell type. Using this system, we biochemically dissect molecular events and signaling pathways involved in NADPH oxidase assembly and demonstrate specific roles for PKCdelta, PI(3,4)P(2)/PI(3,4,5)P(3), and PI(3)P in the PMA-dependent intracellular activation process. This system should be of great utility for the study of cell signaling events that regulate the intracellular production of reactive oxygen species by neutrophils.
机译:活化的嗜中性粒细胞组装一个NADPH氧化酶复合物,以产生用于杀死微生物的超氧化物。大多数初始氧化酶组装发生在细胞内颗粒上,然后氧化酶向吞噬溶酶体和质膜移动。我们已经开发了一种使用链球菌溶血素-O透化的嗜中性粒细胞的新型检测系统,该系统概括了初始细胞内激活过程,同时保持了这种粒细胞类型的超微结构特征。使用该系统,我们生化分析涉及NADPH氧化酶组装的分子事件和信号传导途径,并证明PKCdelta,PI(3,4)P(2)/ PI(3,4,5)P(3)和PI具有特定作用(3)P在依赖PMA的细胞内激活过程中。该系统对于调节中性粒细胞调节活性氧物质的细胞内产生的细胞信号转导事件的研究应具有很大的实用性。

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