Risteli L, Myllylä R, Kivirikko K I
Eur J Biochem. 1976 Aug 1;67(1):197-202. doi: 10.1111/j.1432-1033.1976.tb10649.x.
Denatured citrate-soluble collagen was coupled to agarose by the cyanogen bromide activation technique, and columns prepared from this material were studied for affinity chromatography of collagen glycosyltransferases. Both collagen glycosyltransferases became bound to the column, the degree of binding and the capacity of the column being higher with the glucosyltransferase than with the galactosyltransferase than with the galactosyltransferase. The addition of Mn2+ enhanced the binding, especially with the glucosyltransferase. The enzymes were eluted from the column with small peptides prepared from collagen, and they were separated from the peptides by gel filtration. With this procedure a collagen glucosyltransferase purification of about 5000-fold and a collagen galactosyltransferase purification of about 1000-fold was obtained from chick embryo extract by relatively simple steps.
通过溴化氰活化技术将变性的柠檬酸盐可溶性胶原蛋白与琼脂糖偶联，并研究用这种材料制备的柱子用于胶原蛋白糖基转移酶的亲和色谱分析。两种胶原蛋白糖基转移酶都能与柱子结合，葡糖基转移酶的结合程度和柱子容量比半乳糖基转移酶更高。添加Mn2+可增强结合，尤其是对葡糖基转移酶。用从胶原蛋白制备的小肽将酶从柱子上洗脱下来，并通过凝胶过滤将它们与肽分离。通过相对简单的步骤，从鸡胚提取物中获得了约5000倍的胶原蛋白葡糖基转移酶纯化倍数和约1000倍的胶原蛋白半乳糖基转移酶纯化倍数。
