Guan J L, Machamer C E, Rose J K
Cell. 1985 Sep;42(2):489-96. doi: 10.1016/0092-8674(85)90106-0.
We have previously described the construction and expression of a hybrid gene encoding a membrane-anchored form of rat growth hormone. This protein is anchored in cellular membranes by a carboxy-terminal extension composed of the transmembrane and cytoplasmic domains of the vesicular stomatitis virus glycoprotein. The protein is transported efficiently to the Golgi apparatus but not to the cell surface. To examine the possibility that N-linked glycosylation might be required for protein transport to the cell surface, we created two mutant proteins (using in vitro mutagenesis) in which single amino acids at two random sites in anchored growth hormone were changed to generate consensus sequences required for addition of N-linked carbohydrate. These mutant proteins, and a protein with both glycosylation sites, were glycosylated and were transported to the cell surface. These results suggest that N-linked glycosylation can serve as a signal for protein transport to the cell surface.
我们之前描述过一种编码大鼠生长激素膜锚定形式的杂交基因的构建与表达。该蛋白通过由水泡性口炎病毒糖蛋白的跨膜和胞质结构域组成的羧基末端延伸锚定在细胞膜中。该蛋白能有效地转运至高尔基体，但不能转运至细胞表面。为了研究N-连接糖基化对于蛋白转运至细胞表面是否可能是必需的，我们（利用体外诱变）创建了两种突变蛋白，其中锚定生长激素中两个随机位点的单个氨基酸被改变，以产生添加N-连接碳水化合物所需的共有序列。这些突变蛋白以及具有两个糖基化位点的蛋白都进行了糖基化，并被转运至细胞表面。这些结果表明，N-连接糖基化可以作为蛋白转运至细胞表面的信号。
