Akhrem A A, Gilevich S N, Shkumatov V M, Chashchin V L
Biomed Biochim Acta. 1984;43(2):165-77.
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 modification of free carboxyl groups by histamine. Dissociation constants have been determined for complexes of adrenodoxin, hepatoredoxin , cytochrome b5 heme-containing tryptic fragment and myoglobin with cytochrome c preparations immobilized via lysine residues (cytochrome c-Sepharose I) or additional imidazole groups (cytochrome c-Sepharose II). The latter adsorbent possesses a 2-3 times higher affinity to adrenodoxin and hepatoredoxin than the former. The parameters of interaction with cytochrome c-Sepharose II constitute for the proteins studied the following sequence: adrenodoxin (the highest affinity) greater than or equal to hepatoredoxin greater than cytochrome b4 heme- containing tryptic fragment greater than myoglobin. The efficiency of cytochrome c-Sepharose II application in the course of adrenodoxin, hepatoredoxin and cytochrome b5 purification, as well as isolation of cytochrome b4 heme-containing tryptic fragment has been shown.
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含血红素胰蛋白酶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（细胞色素c-琼脂糖I）或额外的咪唑基团（细胞色素c-琼脂糖II）形成的复合物的解离常数。后一种吸附剂对肾上腺皮质铁氧还蛋白和肝铁氧还蛋白的亲和力比前一种高2至3倍。与细胞色素c-琼脂糖II相互作用的参数对于所研究的蛋白质构成以下顺序：肾上腺皮质铁氧还蛋白（亲和力最高）≥肝铁氧还蛋白＞细胞色素b4含血红素胰蛋白酶片段＞肌红蛋白。已证明细胞色素c-琼脂糖II在肾上腺皮质铁氧还蛋白、肝铁氧还蛋白和细胞色素b5纯化过程中以及分离细胞色素b4含血红素胰蛋白酶片段方面的应用效率。
