SNAP Prevents Mg-ATP-induced Release of N-Ethylmaleimide-sensitive Factor from the Golgi Apparatus in Digitonin-permeabilized PC12 Cells (*)
SNAP Prevents Mg-ATP-induced Release of N-Ethylmaleimide-sensitive Factor from the Golgi Apparatus in Digitonin-permeabilized PC12 Cells (*)
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SNAP 可防止 Mg-ATP 诱导的洋地黄皂苷透化 PC12 细胞中 N-乙基马来酰亚胺敏感因子从高尔基体释放 (*)
DOI：
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发表时间：
1996
影响因子：
4.8
通讯作者：
S. Mizushima
中科院分区：
文献类型：
--
作者：
M. Tagaya;Akiko Furuno;S. Mizushima
N-乙基马来酰亚胺敏感因子（NSF），这是参与蛋白质转运的多个步骤，在体外孵育与Mg-ATP从高尔基体膜释放。然而，一些证据表明，NSF与膜相关，尽管体内存在Mg和ATP。我们使用毛地黄皂苷透化的PC 12细胞来研究NSF与膜的关联机制。在PC 12细胞中，免疫反应NSF观察到核膜，高尔基体，神经生长锥，突触囊泡集中。与高尔基体相关的NSF在与Mg-ATP孵育时释放，而在核膜和神经元生长锥中的NSF在相同的处理中不释放。胞质溶胶的加入阻断了Mg-ATP诱导的NSF从高尔基体的释放。色谱分析显示，阻止NSF从高尔基体释放的因子在与可溶性NSF附着蛋白（SNAP）相同的位置洗脱。纯化的His标记的α-SNAP表现出这种活性。His标记的α-SNAP也阻止Mg-ATP诱导的NSF从分离的高尔基体膜释放。
The N-ethylmaleimide-sensitive factor (NSF), which is involved in the multisteps of protein transport, is released from Golgi membranes on in vitro incubation with Mg-ATP. However, several lines of evidence suggest that NSF is associated with membranes in spite of the presence of Mg and ATP in vivo. We have used digitonin-permeabilized PC12 cells to investigate the mechanism underlying the association of NSF with membranes. In PC12 cells, immunoreactivity for NSF was observed in the nuclear membranes, the Golgi apparatus, and neuronal growth cones, where synaptic vesicles are concentrated. NSF associated with the Golgi apparatus was released on incubation with Mg-ATP, whereas NSF in the nuclear membranes and neuronal growth cones was not released on the same treatment. The addition of cytosol blocked the Mg-ATP-induced release of NSF from the Golgi apparatus. Chromatographic analyses revealed that the factor(s) that prevents NSF release from the Golgi apparatus was eluted at the same position as the soluble NSF attachment proteins (SNAPs). Purified His-tagged α-SNAP exhibited such activity. His-tagged α-SNAP also prevented the Mg-ATP-induced release of NSF from isolated Golgi membranes.
DOI：
10.1073/pnas.82.12.4137
发表时间：
1985-01-01
影响因子：
11.1
作者：
JAHN, R;SCHIEBLER, W;GREENGARD, P
通讯作者：
GREENGARD, P
DOI：
10.1073/pnas.85.21.7852
发表时间：
1988-11-01
影响因子：
11.1
作者：
BLOCK, MR;GLICK, BS;ROTHMAN, JE
通讯作者：
ROTHMAN, JE
纯化囊泡运输所需的三种相关外周膜蛋白。
DOI：
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发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Clary,DO;Rothman,JE
通讯作者：
Rothman,JE