Wallis M, Davies R V, Daniels M
Biochem J. 1978 Mar 1;169(3):669-76. doi: 10.1042/bj1690669.
Rat somatotropin (growth hormone) was labelled biosynthetically by incubating anterior pituitary lobes with radioactive amino acids for 24 h in a simple buffered salts medium containing glucose. The labelled hormone was isolated by preparative polyacrylamide-gel electrophoresis or by chromatography on Sephadex G-100 and then DEAE-cellulose. The labelled material was pure by several criteria and cross-reacted immunologically with unlabelled rat somatotropin. When a mixture of 14C-labelled amino acids was used for labelling the protein, label could be introduced into these same amino acids of somatotropin, though relative specific radioactivities varied considerably. Somatotropin labelled by the procedures described in the present paper was suitable for structural studies and could be used for a variety of other biochemical experiments.
通过在含有葡萄糖的简单缓冲盐培养基中,将垂体前叶与放射性氨基酸一起孵育24小时,对大鼠生长激素进行生物合成标记。标记的激素通过制备性聚丙烯酰胺凝胶电泳或在葡聚糖G - 100上进行色谱分离,然后再通过二乙氨基乙基纤维素柱色谱进行分离。通过多项标准判断,标记物是纯的,并且能与未标记的大鼠生长激素发生免疫交叉反应。当使用14C标记氨基酸混合物对蛋白质进行标记时,标记可以引入到生长激素的相同氨基酸中,尽管相对比放射性差异很大。用本文所述方法标记的生长激素适用于结构研究,并可用于各种其他生化实验。