REGULATION OF GLYCOPROTEIN BIOSYNTHESIS

拟议的工作涉及开发和使用一种测定方法， 磷酸长羟乙酯，一种参与糖蛋白生物合成的磷脂。 的 测定是基于使用DEAE纤维素纯化磷酸长羟乙酯 和衍生化前的二维薄层色谱法， 使用氯甲酸C14-苯基酯定量。 还提出了一项研究， 在肝细胞中，C14-乙酸酯掺入胆固醇， 将监测磷酸长羟乙酯作为HMG-CoA水平的函数 还原酶，这将在不同程度上抑制使用特定的 抗生素康帕汀 这些研究将提供有关这些因素的信息， 其控制着磷酸长链酯生物合成的速率。 最后一个协议 概述了UDPG 1cNAc-1-P转移酶的亲和层析， 与不溶性树脂结合的特异性抑制剂衣霉素。

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.