Respiratory burst enzyme in human neutrophils. Evidence for multiple mechanisms of activation.
Respiratory burst enzyme in human neutrophils. Evidence for multiple mechanisms of activation.
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人类中性粒细胞中的呼吸爆发酶。
DOI：
10.1172/jci110087
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发表时间：
1981
期刊：
影响因子：
--
通讯作者：
JohnstonJr,RB
中科院分区：
文献类型：
--
作者：
McPhail,LC;Henson,PM;JohnstonJr,RB
发现非穿透性蛋白质灭活剂对-重氮苯磺酸（DASA）对人中性粒细胞表面的改变可阻止某些刺激引起的呼吸爆发的激活，但对其他刺激不起作用。DASA预处理可抑制豆豆蛋白A或趋化肽甲酰基-蛋氨酸-乙酰氨基-苯丙氨酸FMLP刺激超氧阴离子（O2-）的产生，而肉豆酸酯佛波酯（PMA）、氟化钠刺激O2-的产生。或离子载体A23187不受DASA的抑制。DASA预处理抑制FMLP刺激的氧摄取，但不抑制PMA刺激的氧摄取。DASA可重复性地抑制两种已知表面酶Mg++- atp酶和碱性磷酸酶的活性，分别为45-55%和60-70%。DASA对O2生成的抑制似乎不是由干扰受影响刺激的结合引起的，因为DASA预处理不会抑制豆豆蛋白A或FMLP诱导的溶酶体酶溶菌酶和髓过氧化物酶的释放。来自中性粒细胞的富膜颗粒组分已被证明含有nadph依赖的氧化活性，这可能是完整细胞吞噬相关呼吸爆发的原因。pma活化酶易受直接暴露于DASA颗粒组分的抑制。这些发现表明，在人类中性粒细胞中存在不止一种激活呼吸爆发氧化酶的机制，并且中性粒细胞至少具有一种非外切酶的氧化酶。
Alteration of the surface of human neutrophils with the nonpenetrating, protein-inactivating agent p-diazobenzenesulfonic acid (DASA) was found to prevent activation of the respiratory burst by some stimuli, but not others. Production of superoxide anion (O2-) stimulated by concanavalin A or the chemotactic peptide formyl-methionyl-leucyl-phenylalanine FMLP was inhibited by DASA pretreatment, whereas O2- production stimulated by phorbol myristate acetate (PMA), sodium fluoride. or the ionophore A23187 was not inhibited by DASA. Pretreatment with DASA inhibited oxygen uptake stimulated by FMLP, but not oxygen uptake stimulated by PMA. DASA reproducibly inhibited activities of two known surface enzymes Mg++-ATPase and alkaline phosphatase, by 45-55% and 60-70%, respectively. The inhibition by DASA of O2- production did not appear to be caused by interference with binding of the affected stimuli, since pretreatment with DASA did not inhibit release of the lysosomal enzymes lysozyme and myeloperoxidase induced by concanavalin A or FMLP. Membrane-rich particulate fractions from neutrophils have been shown to contain NADPH-dependent oxidative activity that is presumably responsible for the phagocytosis-associated respiratory burst of intact cells. The PMA-activated enzyme was susceptible to inhibition of directly exposed to DASA in this particulate fraction. These findings suggest that more than one mechanism exists for activation of the respiratory burst oxidase in human neutrophils, and that the neutrophil possesses at least one oxidase that is not an ectoenzyme.