Vesicular and nonvesicular transport of ceramide from ER to the Golgi apparatus in yeast.

Vesicular and nonvesicular transport of ceramide from ER to the Golgi apparatus in yeast.
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DOI:
10.1083/jcb.200105033
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发表时间:
2001-12-10
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Riezman H
Riezman H
中科院分区:
其他
文献类型:
--
作者:
Funato K;Riezman H

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脂质的运输和分选必须以特定的机制发生,因为细胞内细胞器的膜在脂质组成上不同,即使大多数脂质生物合成开始于ER。在酵母中,神经酰胺在内质网中合成,并转移到高尔基体,在那里形成肌醇磷酸神经酰胺(IPC)。这两个事实意味着,神经酰胺可以被运送到高尔基体独立的囊泡交通,因为IPC的合成仍然继续时,囊泡运输被阻断在sec突变体。完整细胞中的非囊泡IPC合成不受ATP耗竭的影响。使用体外试验,重建神经酰胺的非囊泡途径的运输,我们发现,运输是温度和胞质溶胶依赖性,但能量独立。ER和高尔基体组分在4°C下一起预孵育,其中神经酰胺转运不发生,使得转运反应膜浓度无关,提供ER-高尔基体膜接触刺激神经酰胺转运的生化证据。ER-高尔基体接触建立后,需要一种胞浆蛋白酶敏感因子。
Transport and sorting of lipids must occur with specific mechanisms because the membranes of intracellular organelles differ in lipid composition even though most lipid biosynthesis begins in the ER. In yeast, ceramide is synthesized in the ER and transferred to the Golgi apparatus where inositolphosphorylceramide (IPC) is formed. These two facts imply that ceramide can be transported to the Golgi independent of vesicular traffic because IPC synthesis still continues when vesicular transport is blocked in sec mutants. Nonvesicular IPC synthesis in intact cells is not affected by ATP depletion. Using an in vitro assay that reconstitutes the nonvesicular pathway for transport of ceramide, we found that transport is temperature and cytosol dependent but energy independent. Preincubation of ER and Golgi fractions together at 4°C, where ceramide transport does not occur, rendered the transport reaction membrane concentration independent, providing biochemical evidence that ER-Golgi membrane contacts stimulate ceramide transport. A cytosolic protease-sensitive factor is required after establishment of ER-Golgi contacts.
将完整的GPI锚固固定在蛋白质上需要酵母GAA1P。
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