Shkumatov V M, Gilevich S N, Chashchin V L, Akhrem A A
Bioorg Khim. 1983 Sep;9(9):1237-47.
Preparations of horse heart cytochrome c have been obtained immobilized on Sepharose derivatives via lysine epsilon-amino groups, carboxyl groups of aspartic and glutamic acid residues, methionine and histidine residues as well as imidazole groups additionally introduced by means of chemical modification of free carboxyl groups by histamine. Dissociation constants have been determined for complexes of adrenodoxin, hepatoredoxin, cytochrome b5 heme-containing fragment and myoglobin with preparations of cytochrome c immobilized via lysine residues (adsorbent I) or additionally introduced imidazole groups (adsorbent II). The latter is found to possess a 2-3 times greater affinity for adrenodoxin and hepatoredoxin than the former. The affinity of the proteins studied for the adsorbent II constitutes the following sequence: adrenodoxin greater than or equal to hepatoredoxin greater than cytochrome b5 heme-containing fragment greater than myoglobin. The adsorbent II is shown to be effective when used for purification of hepatoredoxin, adrenodoxin, cytochrome b5 and isolation of cytochrome b5 heme-containing fragment.
已获得通过赖氨酸ε-氨基、天冬氨酸和谷氨酸残基的羧基、甲硫氨酸和组氨酸残基以及通过组胺对游离羧基进行化学修饰而额外引入的咪唑基团固定在琼脂糖衍生物上的马心脏细胞色素c制剂。已测定了肾上腺皮质铁氧还蛋白、肝铁氧还蛋白、细胞色素b5含血红素片段和肌红蛋白与通过赖氨酸残基固定的细胞色素c制剂（吸附剂I）或额外引入的咪唑基团（吸附剂II）形成的复合物的解离常数。发现后者对肾上腺皮质铁氧还蛋白和肝铁氧还蛋白的亲和力比前者高2至3倍。所研究的蛋白质对吸附剂II的亲和力构成以下顺序：肾上腺皮质铁氧还蛋白≥肝铁氧还蛋白＞细胞色素b5含血红素片段＞肌红蛋白。结果表明，吸附剂II用于纯化肝铁氧还蛋白、肾上腺皮质铁氧还蛋白、细胞色素b5以及分离细胞色素b5含血红素片段时是有效的。
