猪肝线粒体中的电子传递黄素蛋白。一种简单的纯化方法及对某些分子特性的重新评估。
Electron transfer flavoprotein from pig liver mitochondria. A simple purification and re-evaluation of some of the molecular properties.
作者信息
Husain M, Steenkamp D J
出版信息
Biochem J. 1983 Feb 1;209(2):541-5. doi: 10.1042/bj2090541.
Abstract
Electron transfer flavoprotein (ETF) from pig liver mitochondria has been purified to homogeneity by a three-step procedure with approx. 10-fold higher yields than previously reported. The purified ETF exhibits an absorption coefficient for the bound FAD of 13.5 mM-1.cm-1 at 436 nm and an isoelectric point of 6.75. Gel filtration, sodium dodecyl sulphate gel electrophoresis and flavin analysis indicate that pig liver ETF is a dimer, composed of non-identical subunits (Mr 38 000 and 32 000) with only one FAD/dimer. Anaerobic reduction by dithionite produces anionic flavin semiquinone as a stable intermediate and establishes the flavin to be the only redox-active chromophore in ETF.
摘要
通过三步纯化程序，猪肝线粒体中的电子传递黄素蛋白（ETF）已被纯化至均一状态，产率比之前报道的高约10倍。纯化后的ETF在436nm处结合FAD的吸收系数为13.5 mM-1.cm-1，等电点为6.75。凝胶过滤、十二烷基硫酸钠凝胶电泳和黄素分析表明，猪肝ETF是一种二聚体，由不相同的亚基（Mr 38 000和32 000）组成，每个二聚体仅含一个FAD。连二亚硫酸盐的厌氧还原产生阴离子黄素半醌作为稳定中间体，并确定黄素是ETF中唯一具有氧化还原活性的发色团。
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