Safi A, Saunier M, Gastin I, Alibada Y, Dugue B, Gueant J L
Laboratory of Cellular and Molecular Biology in Nutrition, Medical Faculty, University of Nancy, France.
J Chromatogr B Biomed Appl. 1995 Feb 3;664(1):253-9. doi: 10.1016/0378-4347(94)00426-6.
We have identified a soluble receptor for intrinsic factor (IF) in human urine. The purification of this protein by affinity chromatography required a preliminary purification of IF from hog pyloric mucosal extract. This was achieved by thermolabile cobalamin-ethanol-aminohexane Sepharose affinity chromatography with a 133-fold purification, a yield of 45% and a specific binding activity of 15720 pmol/mg protein. The purified Cbl-IF complex was coupled to epoxy-Sepharose with a yield of 23.8% and a specific activity of 1.2 nmol per mol of gel. The soluble IF receptor was purified form 200 ml of urine concentrate of pregnant women. Desorption was performed at pH 5.0 and in the presence of 5 mM EDTA. The soluble IF receptor was purified 17,200-fold with a yield of 52% and a IF binding capacity of 3260 pmol per mg of protein. A single protein with a Mr of 70,000 was found in silver-stained SDS-PAGE.
我们在人尿中鉴定出一种内因子（IF）的可溶性受体。通过亲和层析法纯化这种蛋白质需要先从猪幽门黏膜提取物中纯化IF。这是通过热不稳定钴胺素 - 乙醇 - 氨基己烷琼脂糖亲和层析实现的，纯化倍数为133倍，产率为45%，比结合活性为15720 pmol/mg蛋白质。纯化后的钴胺素 - IF复合物与环氧琼脂糖偶联，产率为23.8%，每摩尔凝胶的比活性为1.2 nmol。可溶性IF受体从200 ml孕妇尿液浓缩物中纯化得到。在pH 5.0和5 mM EDTA存在的条件下进行解吸。可溶性IF受体纯化了17200倍，产率为52%，每毫克蛋白质的IF结合能力为3260 pmol。在银染的SDS - PAGE中发现了一种分子量为70000的单一蛋白质。
