Bellavite P, Jones O T, Cross A R, Papini E, Rossi F
Biochem J. 1984 Nov 1;223(3):639-48. doi: 10.1042/bj2230639.
The superoxide (O2.-)-forming enzyme NADPH oxidase from pig neutrophils was solubilized and partially purified by gel-filtration chromatography. The purification procedure allowed the separation of NADPH oxidase activity from NADH-dependent cytochrome c reductase and 2,6-dichlorophenol-indophenol reductase activities. O2.-forming activity was co-purified with cytochrome b-245 and was associated with phospholipids. However, active fractions endowed with cytochrome b were devoid of ubiquinone and contained only little FAD. The cytochrome b/FAD ratio was 1.13:1 in the crude solubilized extract and increased to 18.95:1 in the partially purified preparations. Most of FAD was associated with fractions containing NADH-dependent oxidoreductases. These results are consistent with the postulated role of cytochrome b in O2.-formation by neutrophil NADPH oxidase, but raise doubts about the participation of flavoproteins in this enzyme activity.
通过凝胶过滤色谱法溶解并部分纯化了来自猪中性粒细胞的超氧化物（O2.-）生成酶NADPH氧化酶。纯化过程使得NADPH氧化酶活性与NADH依赖性细胞色素c还原酶及2,6 - 二氯酚靛酚还原酶活性得以分离。O2.-生成活性与细胞色素b - 245共同纯化，并与磷脂相关。然而，具有细胞色素b的活性级分不含泛醌，仅含有少量FAD。在粗溶解提取物中细胞色素b/FAD比率为1.13:1，在部分纯化制剂中增至18.95:1。大部分FAD与含有NADH依赖性氧化还原酶的级分相关。这些结果与细胞色素b在中性粒细胞NADPH氧化酶形成O2.-过程中所假定的作用一致，但对黄素蛋白参与该酶活性提出了疑问。
