INTRACELLULAR-TRANSPORT OF SECRETORY AND MEMBRANE-PROTEINS IN HEPATOMA-CELLS INFECTED BY VESICULAR STOMATITIS-VIRUS
INTRACELLULAR-TRANSPORT OF SECRETORY AND MEMBRANE-PROTEINS IN HEPATOMA-CELLS INFECTED BY VESICULAR STOMATITIS-VIRUS
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DOI：
10.1016/0092-8674(80)90547-4
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发表时间：
1980-01-01
期刊：
影响因子：
64.5
通讯作者：
LODISH, HF
中科院分区：
文献类型：
--
作者：
STROUS, JAM;LODISH, HF
用RNA复制和转录缺陷的水泡性口炎病毒（VSV）的温度敏感突变体感染分泌几种血清蛋白的大鼠肝癌细胞。这些感染的细胞在39.5 ℃下生长。C，继续正常分泌白蛋白和转铁蛋白，因此可以在同一细胞中研究分泌蛋白和膜蛋白（包括VSV G蛋白）的合成和成熟。血清蛋白不分泌的细胞在相同的速度，一半的新合成的白蛋白出现在培养基中23分钟后，而一半的糖蛋白转铁蛋白分泌后，只有约40分钟。两个膜蛋白，VSV G糖蛋白和高分子量肝癌表面蛋白，检测在细胞表面约23分钟后，其合成。分泌和膜糖蛋白之间的外化速率的差异是由于从粗面内质网（ER）进入或通过高尔基体区域的运输速率的差异，如通过Asn连接的寡糖成熟所需的时间来判断的。未糖基化的VSV G不会到达细胞表面;未糖基化的转铁蛋白以与糖基化物质相同的速率分泌。因此，合成或Asn连接的寡糖链的存在都不是转铁蛋白分泌速率的决定因素。单价离子载体莫能菌素抑制血清蛋白的分泌和膜蛋白向细胞表面的迁移。转铁蛋白的细胞内转运在分子仍然对内切-β-转铁蛋白敏感的阶段被阻断。N-乙酰氨基葡萄糖苷酶H; VSV G达到对该酶具有抗性的阶段。因此，对于膜和分泌蛋白和糖蛋白，细胞内途径的至少一部分可能是不同的。
Rat hepatoma cells which secrete several serum proteins were infected with a temperature-sensitive mutant of vesicular stomatitis virus (VSV) which is defective in RNA replication and transcription. These infected cells, grown at 39.5.degree. C, continue normal secretion of albumin and transferrin, so that synthesis and maturation of secretory and membrane proteins, including the VSV G protein, can be studied in the same cells. Serum proteins are not secreted from the cells at the same rate; half of the newly synthesized albumin appears in the medium after 23 min, while half of the glycoprotein transferrin is secreted only after about 40 min. Two membrane proteins, the VSV G glycoprotein and a high MW hepatoma surface protein, are detected at the cell surface approximately 23 min after their synthesis. The differences in the rate of externalization among the secretory and membrane glycoproteins are due to differences in the rate of transport from the rough endoplasmic reticulum (ER) into or through Golgi region, as judged by the time required for maturation of the Asn-linked oligosaccharides. Unglycosylated VSV G does not reach the cell surface; unglycosylated transferrin is secreted at the same rate as the glycosylated species. Thus neither the synthesis nor the presence of the Asn-linked oligosaccharide chain are determinants in the rate of transferrin secretion. The monovalent ionophore monensin inhibits secretion of serum proteins and migration of membrane proteins to the cell surface. The intracellular transport of transferrin is blocked in a stage where the molecule is still sensitive to endo-.beta.-N-acetylglucosaminidase H; VSV G reaches a stage in which it is resistant to the enzyme. Thus, at least part of the intracellular pathway may be different for membrane and secretory proteins and glycoproteins.