Borregaard N, Tauber A I
J Biol Chem. 1984 Jan 10;259(1):47-52.
Human neutrophils were fractionated by nitrogen cavitation and Percoll density centrifugation, and the subcellular localization of FAD-flavoprotein, b-cytochrome, NADH-cytochrome b5 reductase, and NADPH-dependent cytochrome c reductase were determined in normal cells, cells from two patients with chronic granulomatous disease (CGD), and normal cells that had been stimulated with phorbol myristate acetate. In normal cells, a FAD-flavoprotein is found in a 1:2 molar ratio, with cytochrome b in the fractions containing the specific granules. Triton X-114 phase distribution indicates that the b-cytochrome but not the b-cytochrome-associated flavoprotein is an integral membrane protein. 80% of this flavoprotein, as well as all the b-cytochrome, was absent in these fractions from 2 CGD patients, although these patients had normal quantities of FAD in the fractions containing plasma membranes and cytosol. During stimulation the b-cytochrome-associated flavoprotein of the granules translocates with the b-cytochrome to the plasma membrane where NADPH oxidase is localized. Definition of the role of these NADPH oxidase constituents may provide a molecular description of the normal neutrophil respiratory burst and the molecular defect(s) in CGD.
通过氮气空化和Percoll密度离心法对人中性粒细胞进行分级分离，并在正常细胞、两名慢性肉芽肿病（CGD）患者的细胞以及用佛波酯肉豆蔻酸酯刺激过的正常细胞中，测定黄素腺嘌呤二核苷酸（FAD）黄素蛋白、b型细胞色素、NADH - 细胞色素b5还原酶和NADPH依赖性细胞色素c还原酶的亚细胞定位。在正常细胞中，FAD黄素蛋白与细胞色素b以1:2的摩尔比存在于含有特异性颗粒的组分中。Triton X - 114相分配表明，b型细胞色素而非与b型细胞色素相关的黄素蛋白是一种整合膜蛋白。尽管这两名CGD患者含有质膜和胞质溶胶的组分中FAD含量正常，但在这些组分中，这两名患者的80%的这种黄素蛋白以及所有的b型细胞色素均缺失。在刺激过程中，颗粒中与b型细胞色素相关的黄素蛋白与b型细胞色素一起转移至质膜，而NADPH氧化酶定位于质膜。确定这些NADPH氧化酶成分的作用可能会为正常中性粒细胞呼吸爆发以及CGD中的分子缺陷提供分子层面的描述。
