Membrane glycoprotein folding, oligomerization and intracellular transport: effects of dithiothreitol in living cells.
Membrane glycoprotein folding, oligomerization and intracellular transport: effects of dithiothreitol in living cells.
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膜糖蛋白折叠、寡聚化和细胞内转运：二硫苏糖醇对活细胞的影响。
DOI：
10.1002/j.1460-2075.1993.tb05863.x
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Helenius,A
中科院分区：
文献类型：
--
作者：
Tatu,U;Braakman,I;Helenius,A
以流感病毒血凝素（HA0）和水泡性口炎病毒G蛋白为模型蛋白，分析了二硫苏糖醇（DTT）对活细胞分泌途径中糖蛋白构象成熟和转运的影响。虽然DTT导致内质网（ER）中折叠中间体和错误折叠蛋白质的减少，但它并不影响已经获得成熟三聚体构象的分子，无论是在ER中还是在其他地方。因此，向DTT耐药的转化是高尔基体前事件。折叠中间体的减少依赖于ER的完整性和代谢能量，这表明DTT和ER折叠因子之间的协同性。DTT不抑制大多数细胞功能，包括分泌途径内的ATP合成和蛋白质转运。结果表明，DTT是分析含二硫键蛋白质折叠和区室分布的有效工具。
Using influenza hemagglutinin (HA0) and vesicular stomatitis virus G protein as model proteins, we have analyzed the effects of dithiothreitol (DTT) on conformational maturation and transport of glycoproteins in the secretory pathway of living cells. While DTT caused reduction of folding intermediates and misfolded proteins in the endoplasmic reticulum (ER), it did not affect molecules that had already acquired a mature trimeric conformation, whether present in the ER or elsewhere. The conversion to DTT resistance was therefore a pre‐Golgi event. Reduction of folding intermediates was dependent on the intactness of the ER and on metabolic energy, suggesting cooperativity between DTT and ER folding factors. DTT did not inhibit most cellular functions, including ATP synthesis and protein transport within the secretory pathway. The results established DTT as an effective tool for analyzing the folding and compartmental distribution of proteins with disulfide bonds.