SEMI-INTACT CELLS PERMEABLE TO MACROMOLECULES - USE IN RECONSTITUTION OF PROTEIN-TRANSPORT FROM THE ENDOPLASMIC-RETICULUM TO THE GOLGI-COMPLEX
SEMI-INTACT CELLS PERMEABLE TO MACROMOLECULES - USE IN RECONSTITUTION OF PROTEIN-TRANSPORT FROM THE ENDOPLASMIC-RETICULUM TO THE GOLGI-COMPLEX
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DOI：
10.1016/0092-8674(87)90025-0
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发表时间：
1987-08-14
期刊：
影响因子：
64.5
通讯作者：
BALCH, WE
中科院分区：
文献类型：
--
作者：
BECKERS, CJM;KELLER, DS;BALCH, WE
我们引入了一种新方法，可以去除真核细胞的部分质膜以形成半完整的细胞。在制备过程中，这些细胞失去了可溶性细胞质内容物，但保留了完整形式的分泌细胞器，例如内质网和高尔基复合体。通过在细胞质和 ATP 存在下孵育，可以使用这些半完整细胞在体外功能性地重建 ER 和高尔基体之间的蛋白质运输。体外ER中水泡性口炎病毒株tsO45 G蛋白的输出对温度敏感，与体内观察到的结果相似。这些细胞允许化学物质和抗体直接进入细胞的细胞质结构域，并且可能是广泛适用的模型系统，用于研究细胞生物学中的广泛问题。
We introduce a new method that removes portions of the plasma membrane of eukaryotic cells to form semi-intact cells. During preparation, these cells lose their soluble cytoplasmic contents, but retain secretory organelles such as the ER and Golgi complex in an intact form. Transport of protein between the ER and Golgi can be functionally reconstituted in vitro using these semi-intact cells by incubation in the presence of cytosol and ATP. Export of the vesicular stomatiatis virus strain tsO45 G protein from the ER in vitro is temperature-sensitive, similar to the result observed in vivo. These cells allow direct access of chemicals and antibodies to the cytoplasmic domain of the cell and may be a widely applicable model system for study of a broad range of problems in cell biology.