REGULATION OF GLYCOPROTEIN BIOSYNTHESIS

拟议的工作涉及开发和使用一种用于 磷脂，一种参与糖蛋白生物合成的磷脂。这个 测定是基于使用DEAE纤维素纯化磷酸苯二醇酯 以及衍生化前的二维薄层层析和 使用C14-苯基氯甲酸酯进行定量。还提出了一项使用RAT进行的研究 在肝细胞中，C14-乙酸酯掺入胆固醇和 将监测作为HMG-CoA水平的函数的磷酸二氢呋喃 还原酶，它将被不同程度地抑制使用特定的 抗生素紧凑剂。这样的研究将提供有关这些因素的信息 它们控制着磷酸二氢呋喃的生物合成速率。最后是一项协议 UDPG1cNAc-1-P转移酶的亲和层析 结合在不溶树脂上的特定的衣霉素抑制剂。

The work proposed involves the development and employment of an assay for dolichyl phosphate, a phospholipid involved in glycoprotein biosynthesis. The assay is based on the purification of dolichyl phosphate using DEAE cellulose and two-dimensional thin-layer chromatography prior to derivatization and quantitation using C14-phenyl chloroformate. Also proposed is a study using rat liver cells in which the incorporation of C14-acetate into cholesterol and dolichyl phosphate will be monitored as a function of the level of HMG-CoA reductase, which will be inhibited to varying degrees using the specific antibiotic compactin. Such studies will provide information on the factors which control the rate of dolichyl phosphate biosynthesis. Finally a protocol is outlined for affinity chromatography of UDPG1cNAc-1-P transferase using the specific inhibitor tunicamycin bound to an insoluble resin.