Shkumatov V M, Gilevich S N, Chashchin V L, Akhrem A A
Bioorg Khim. 1983 Sep;9(9):1231-6.
A technique for the isolation of highly purified hepatoredoxin involving the DE-32 cellulose chromatography of post-mitochondrial supernatant, ammonium-sulfate fractionation, Sephadex G-75 gel chromatography, 1-amino-2-hydroxypropyl-Sepharose ion-exchange chromatography and cytochrome-c-Sepharose affinity chromatography is described. The protein was purified 160-fold with a yield of 19%. The synthesis of cytochrome-c-Sepharose was carried out in a way preventing modification of the lysine-containing binding domain of the cytochrome c molecule. To achieve this, free carboxyl groups were modified with histamine to introduce imidazole residues in cytochrome c and the modified protein was immobilized on bromoacetyl-Sepharose.
描述了一种分离高度纯化的肝红素氧化还原蛋白的技术，该技术包括线粒体后上清液的DE-32纤维素色谱法、硫酸铵分级分离、Sephadex G-75凝胶色谱法、1-氨基-2-羟丙基-Sepharose离子交换色谱法和细胞色素c-Sepharose亲和色谱法。该蛋白质被纯化了160倍，产率为19%。细胞色素c-Sepharose的合成是以防止细胞色素c分子含赖氨酸的结合域发生修饰的方式进行的。为实现这一点，用组胺修饰游离羧基以在细胞色素c中引入咪唑残基，然后将修饰后的蛋白质固定在溴乙酰-Sepharose上。
