PASSAGE OF VIRAL MEMBRANE-PROTEINS THROUGH THE GOLGI-COMPLEX
PASSAGE OF VIRAL MEMBRANE-PROTEINS THROUGH THE GOLGI-COMPLEX
复制标题
DOI：
10.1016/0022-2836(81)90122-4
复制
发表时间：
1981-01-01
影响因子：
5.6
通讯作者：
WARREN, G
中科院分区：
文献类型：
--
作者：
GREEN, J;GRIFFITHS, G;WARREN, G
用放线菌酮处理感染塞姆利基森林病毒的BHK-21细胞，以阻止病毒膜蛋白的进一步合成，但不阻止病毒膜蛋白在细胞内的转运。然后使用间接标记铁蛋白或Au的特定抗体将这些蛋白质定位在薄的冰冻切片中。显微照片上标记的定量显示，刺突蛋白从粗面内质网移动并通过高尔基体堆叠。尖峰蛋白花费了.apprx。15分钟。在这些细胞内细胞器中，它们的最终目的地是质膜。平行的生化研究表明，病毒刺突蛋白上的大多数简单低聚糖在这些膜蛋白通过高尔基堆叠的同时被修饰成复杂的形式。细胞分级研究揭示了相同的模式：蛋白质通过根据半乳糖转移酶含量分离的囊泡部分从粗面内质网进入质膜。该组分至少部分来源于BHK细胞中的高尔基复合体，这一独立证据表明，它与一种针对大鼠肝脏高尔基体膜的抗体发生特异性反应。
BHK-21 cells [baby hamster kidney] infected with Semliki Forest virus, were treated with cycloheximide to stop further synthesis but not intracellular transport of the viral membrane proteins. These proteins were then localized in thin, frozen sections using specific antibodies labeled indirectly with ferritin or Au. Quantitation of the labeling on micrographs showed the movement of spike proteins from the rough endoplasmic reticulum and through the Golgi stacks. The spike proteins spent .apprx. 15 min. in each of these intracellular organelles; their final destination was the plasma membrane. Parallel biochemical studies showed that most of the simple oligosaccharides on the viral spike proteins were modified to the complex form at the same time as these membrane proteins were passing through the Golgi stacks. Cell fractionation studies revealed the same pattern; the proteins passed from the rough endoplasmic reticulum to the plasma membrane via a vesicle fraction isolated according to its content of galactosyl transferase. Independent evidence that this fraction was derived at least partly from the Golgi complex in BHK cells was obtained by showing that it reacted specifically with an antibody raised to rat liver Golgi membranes.