MANIPULATING DISULFIDE BOND FORMATION AND PROTEIN FOLDING IN THE ENDOPLASMIC-RETICULUM
MANIPULATING DISULFIDE BOND FORMATION AND PROTEIN FOLDING IN THE ENDOPLASMIC-RETICULUM
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DOI：
10.1002/j.1460-2075.1992.tb05223.x
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发表时间：
1992-05-01
期刊：
影响因子：
11.4
通讯作者：
HELENIUS, A
中科院分区：
文献类型：
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作者：
BRAAKMAN, I;HELENIUS, J;HELENIUS, A
在活细胞培养液中加入还原剂二硫苏糖醇(DTT)可阻止新合成的流感血凝素(HA0)中二硫键的形成，并诱导内质网内已氧化的HA0的还原。减少的HA0没有三聚化或离开ER。当DTT被洗掉时，HA0被迅速氧化，正确折叠，三聚，并被输送到高尔基复合体。我们的结论是，添加DTT可以很容易地操纵内质网中的蛋白质折叠和氧化还原条件，而不影响大多数其他细胞功能，减少的流感HA0在很大程度上仍然没有展开，通常发生在新生HAO链上的折叠事件可以在翻译后延迟和诱导，而不会降低效率。
Addition of the reducing agent dithiothreitol (DTT) to the medium of living cells prevented disulfide bond formation in newly synthesized influenza hemagglutinin (HA0) and induced the reduction of already oxidized HA0 inside the ER. The reduced HA0 did not trimerize or leave the ER. When DTT was washed out, HA0 was rapidly oxidized, correctly folded, trimerized and transported to the Golgi complex. We concluded that protein folding and the redox conditions in the ER can be readily manipulated by addition of DTT without affecting most other cellular functions, that the reduced influenza HA0 remains largely unfolded, and that folding events that normally take place on the nascent HAO chains can be delayed and induced post-translationally without loss in efficiency.