Cooke E, Hallett M B
Biochem J. 1985 Dec 1;232(2):323-7. doi: 10.1042/bj2320323.
The role of C-kinase in the triggering of the neutrophil oxidase by two stimuli (latex beads and the chemotactic peptide fMet-Leu-Phe), representative of endocytotic and exocytotic routes of activation, were investigated by using experimental agents that activate, or inhibit C-kinase, in intact cells. The activation by the phagocytotic stimulus latex beads was mimicked by C-kinase activators giving the same characteristic lag (20-30s), followed by a constant oxygen consumption rate with the same maximum rate and affinity for oxygen (Km approx. 13 microM), competed with activation by PMA (4 beta-phorbol 12-myristate 13-acetate) in a simple common-target manner, and was inhibited by retinal, an inhibitor shown to inhibit activation by PMA. In contrast, activation by chemotactic peptide was not mimicked by C-kinase activation alone, chemotactic peptide inducing biphasic oxygen consumption with a Km for oxygen of the second prolonged phase of 3.9 microM, did not compete with activation by PMA, and was not inhibited by retinal. However, PMA and retinal produced slight enhancements of activation by chemotactic peptide and production of monophasic oxygen consumption. It was concluded that C-kinase activation plays a simple central transducing role in activation of the oxidase by latex beads, but that its role in activation by chemotactic peptide is a part of a more complex set of interactions that involve other Ca2+-activated and non-Ca2+-activated processes.
通过使用能在完整细胞中激活或抑制C激酶的实验试剂,研究了C激酶在由两种刺激物(乳胶珠和趋化肽fMet-Leu-Phe)触发中性粒细胞氧化酶过程中的作用,这两种刺激物分别代表内吞和外排激活途径。吞噬刺激物乳胶珠所引发的激活作用,可被C激酶激活剂模拟,呈现相同的特征延迟(20 - 30秒),随后是恒定的耗氧速率,具有相同的最大速率和对氧的亲和力(Km约为13微摩尔),以简单的共同靶点方式与佛波酯(4β-佛波醇12-肉豆蔻酸酯13-乙酸酯)激活作用竞争,并被视黄醛抑制,视黄醛是一种已显示可抑制佛波酯激活作用的抑制剂。相比之下,趋化肽的激活作用不能仅被C激酶激活所模拟,趋化肽诱导双相耗氧,第二个延长阶段的氧Km为3.9微摩尔,不与佛波酯激活作用竞争,也不被视黄醛抑制。然而,佛波酯和视黄醛会轻微增强趋化肽的激活作用以及单相耗氧的产生。得出的结论是,C激酶激活在乳胶珠激活氧化酶过程中起简单的中心转导作用,但它在趋化肽激活过程中的作用是更复杂的一系列相互作用的一部分,这些相互作用涉及其他钙激活和非钙激活过程。