Kolberg J, Michaelsen T E, Jantzen E
Hoppe Seylers Z Physiol Chem. 1983 Feb;364(2):111-7. doi: 10.1515/bchm2.1983.364.1.111.
Human ceruloplasmin was isolated from normal serum by fractional polyethylene glycol precipitation and subsequent ion exchange chromatography. The molecular mass determined by ultracentrifugation was 132 kDa and an optical density ratio (A610/A280) of 0.046 was observed in the preparation. Immunoelectrophoresis, agarose-gel electrophoresis and N-terminal amino-acid sequence analyses showed that the ceruloplasmin preparation was highly purified, while dodecyl sulphate polyacrylamide-gel electrophoresis indicated some degradation of the protein. Affinity chromatography showed that only a fraction of ceruloplasmin was bound to Lens culinaris or Lathyrus odoratus lectin-Sepharose, whereas nearly all the protein was bound to Canavalia ensiformis lectin-Sepharose. The carbohydrate composition of the ceruloplasmin fractions was analysed. It is suggested that fucose might be a determinant for the microheterogeneity in the carbohydrate chains of ceruloplasmin.
通过分级聚乙二醇沉淀和随后的离子交换色谱法从正常血清中分离出人铜蓝蛋白。通过超速离心测定的分子量为132 kDa，并且在制剂中观察到光密度比（A610/A280）为0.046。免疫电泳、琼脂糖凝胶电泳和N端氨基酸序列分析表明，铜蓝蛋白制剂高度纯化，而十二烷基硫酸钠聚丙烯酰胺凝胶电泳表明该蛋白质有一些降解。亲和色谱显示，只有一部分铜蓝蛋白与菜豆或香豌豆凝集素-琼脂糖结合，而几乎所有蛋白质都与刀豆凝集素-琼脂糖结合。分析了铜蓝蛋白组分的碳水化合物组成。有人提出，岩藻糖可能是铜蓝蛋白碳水化合物链中微观异质性的决定因素。
