Activation of the respiratory burst enzyme in human polymorphonuclear leukocytes by chemoattractants and other soluble stimuli. Evidence that the same oxidase is activated by different transductional mechanisms.
Activation of the respiratory burst enzyme in human polymorphonuclear leukocytes by chemoattractants and other soluble stimuli. Evidence that the same oxidase is activated by different transductional mechanisms.
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化学引诱剂和其他可溶性刺激物激活人多形核白细胞中的呼吸爆发酶。
DOI：
10.1172/jci110957
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Snyderman,R
中科院分区：
文献类型：
--
作者：
McPhail,LC;Snyderman,R
趋化因子-受体偶联触发吞噬细胞中的几种生物反应，包括呼吸爆发的激活。先前在完整细胞中的证据表明，化学引诱物刺激呼吸爆发的机制不同于其他可溶性药物，这表明不同的氧化酶负责的可能性。我们现在表明，化学引诱剂N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸和补体第五组分（C5 a）的分裂片段刺激NADPH氧化酶活性，在50，000 g人多形核白细胞（PMN）的颗粒部分中测量。由化学引诱物刺激的氧化酶活性水平是时间和剂量依赖性的，并且在刺激期间需要细胞松弛素B的存在。相反，激活两个非趋化刺激，离子载体A23187和佛波酯肉豆蔻酸酯乙酸酯（PMA），不需要细胞松弛素B。氧化酶激活的时间模式表明，不同的刺激遵循不同的转导途径。化学引诱剂介导的激活是立即的（没有滞后），达到峰值45秒，迅速下降到约50%的最大2分钟。相反，激活A23187或PMA有15-30-s的滞后，增加更慢。A23187的刺激在5 min达到峰值，然后下降。PMA的刺激在20分钟达到平台，并没有下降90分钟。比较Km值的NADPH和NADH得到的Lineweaver-Burk分析的氧化酶活性刺激的N-甲酰-甲硫氨酰-亮氨酰-苯丙氨酸，A23187，和PMA表明，相同的酶被激活的所有刺激。因此，化学引诱物和其他可溶性刺激物似乎激活PMN中相同的呼吸爆发酶，但它们利用不同的转导机制，并且受到不同的调节。
Chemoattractant-receptor coupling triggers several biologic responses in phagocytic cells including activation of the respiratory burst. Prior evidence in intact cells implied that stimulation of the respiratory burst by chemoattractants was by a mechanism different from other soluble agents suggesting the possibility that different oxidative enzymes were responsible. We now show that the chemoattractants N-formyl-methionyl-leucyl-phenylalanine and a split fragment of the fifth component of complement (C5a) stimulate an NADPH oxidase activity, measured in the 50,000-g particulate fraction from human polymorphonuclear leukocytes (PMN). Levels of oxidase activity stimulated by the chemoattractants were both time and dose dependent and required the presence of cytochalasin B during stimulation. In contrast, activation by two nonchemotactic stimuli, the ionophore A23187 and phorbol myristate acetate (PMA), did not require cytochalasin B. Temporal patterns of oxidase activation suggested that different stimuli follow different transductional pathways. Chemoattractant-mediated activation was immediate (no lag); peaked by 45 s and declined rapidly to approximately 50% of maximal by 2 min. In contrast, activation by A23187 or PMA had a 15-30-s lag and increased more slowly. Stimulation by A23187 peaked at 5 min, then declined. Stimulation by PMA plateaued at 20 min and did not decline by 90 min. Comparison of Km values for NADPH and NADH obtained by Lineweaver-Burk analysis of the oxidase activity stimulated by N-formyl-methionyl-leucyl-phenylalanine, A23187, and PMA suggested that the same enzyme was activated by all stimuli. Thus, chemoattractants and other soluble stimuli appear to activate the same respiratory burst enzyme in PMN but they utilize different transductional mechanisms and are regulated differently.
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