Utility of staurosporine in uncovering differences in the signal transduction pathways for superoxide production in neutrophils.
Utility of staurosporine in uncovering differences in the signal transduction pathways for superoxide production in neutrophils.
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星形孢菌素在揭示中性粒细胞超氧化物产生信号转导途径差异方面的用途。
DOI：
10.1016/0167-4889(90)90090-z
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Badwey,JA
中科院分区：
文献类型：
--
作者：
Robinson,JM;Heyworth,PG;Badwey,JA
中性粒细胞在受到phorbol 12-肉豆蔻酸13-乙酸酯（PMA）或fMet-Leu-Phe （fMLP）刺激时，表现出47kda蛋白的强烈磷酸化并释放大量超氧化物（O2−）。蛋白激酶拮抗剂(如200 μM - 1-（5-异喹啉基磺酰基）-2-甲基哌嗪（H-7）；当刺激为PMA时(Badwey， J.A. et al. (1989)；化学，264,14947-14953)。在本文中，我们现在报道，虽然中性粒细胞用15 nM的staurosporine和PMA处理后释放的O2−很少，但在这些化合物存在的情况下，细胞可以通过随后添加fMLP来刺激释放接近正常量的O2−。令人惊讶的是，在fMLP刺激的细胞中，staurosporine （15 nM）至少与H-7一样有效地减少了32p在47 kDa蛋白中的掺入，然而，虽然staurosporine处理的细胞释放了大量的O2−，但H-7处理的细胞却没有。这些数据表明，中性粒细胞中存在一种刺激途径，其中包含一种与PMA刺激时激活的蛋白激酶“不同”的蛋白激酶，这种途径可能使O2 -产生系统在47 kDa蛋白很少或没有磷酸化的情况下发挥作用。他们进一步表明，PMA和fMLP利用的信号转导途径中对H-7敏感的步骤可能不同。
Neutrophils exhibit an intense phosphorylation of a 47 kDa protein and release large quantities of superoxide (O2−) upon stimulation with phorbol 12-myristate 13-acetate (PMA) or fMet-Leu-Phe (fMLP). Antagonists of protein kinases (e.g., 200 μM 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7); 15 nM staurosporine) inhibited these phenomena when the stimulus was PMA (Badwey, J.A. et al. (1989) J. Biol. Chem. 264, 14947–14953). In this paper, we now report that while neutrophils treated with 15 nM staurosporine and PMA release little O2−, cells in the presence of these compounds can be stimulated to release near normal quantities of O2−by the subsequent addition of fMLP. Surprisingly, staurosporine (15 nM) reduced the incorporation of32P into the 47 kDa protein in fMLP stimulated cells at least as effectively as H-7, yet, while the staurosporine treated cells released substantial amounts of O2−, the cells treated with H-7 did not. These data suggest that a stimulatory pathway exists in neutrophils that contains a protein kinase ‘distinct’ from that which is activated when PMA is the stimulus and that this pathway may enable the O2−producing system to become functional with little or no phosphorylation of the 47 kDa protein. They further suggest that the steps which are sensitive to H-7 in the signal-transduction pathways utilized by PMA and fMLP may be different.