Kuwahara T, White R A, Agosin M
Arch Biochem Biophys. 1985 May 15;239(1):18-28. doi: 10.1016/0003-9861(85)90807-0.
A cytosolic flavoprotein enzyme for the protozoan, Trypanosoma cruzi, has been purified essentially to homogeneity by DEAE-cellulose and 2',5'-ADP-agarose column chromatography. The native enzyme has a molecular weight of 100,000 +/- 6,000, is composed of two identical subunits of molecular weight 52,000 +/- 1,000, and contains FAD in the ratio of 1 mol of FAD per mol of enzyme subunit. The enzyme is NADPH-dependent and is capable of reducing cytochrome c, ferricyanide, 2,6-dichloroindophenol, and menadione, but not adrenalin. It does not hydroxylate either sodium salicylate or sodium p-hydroxybenzoate, but N-methylaniline and N,N-dimethylaminobenzaldehyde-supported oxidation of NADPH has been demonstrated. Plots of initial velocity against NADPH concentration give hyperbolic curves with Km values of 6.289 X 10(-5) M. The enzyme is clearly different from the microsomal NADPH-cytochrome c reductase in its intracellular distribution, molecular weight, dimeric nature, presence of only FAD, and activity against secondary and tertiary aromatic amines.
一种针对原生动物克氏锥虫的胞质黄素蛋白酶，通过DEAE-纤维素和2',5'-ADP-琼脂糖柱色谱法已基本纯化至同质。天然酶的分子量为100,000±6,000，由两个分子量为52,000±1,000的相同亚基组成，且每摩尔酶亚基含有1摩尔FAD。该酶依赖NADPH，能够还原细胞色素c、铁氰化物、2,6-二氯靛酚和甲萘醌，但不能还原肾上腺素。它既不能使水杨酸钠或对羟基苯甲酸钠羟化，但已证明N-甲基苯胺和N,N-二甲基氨基苯甲醛支持的NADPH氧化反应。以初始速度对NADPH浓度作图得到双曲线，Km值为6.289×10(-5)M。该酶在细胞内分布、分子量、二聚体性质、仅存在FAD以及对仲胺和叔胺的活性方面明显不同于微粒体NADPH-细胞色素c还原酶。
