Greenberg M E, Edelman G M
Cell. 1983 Jul;33(3):767-79. doi: 10.1016/0092-8674(83)90019-3.
The subcellular localization of the 34 kd protein substrate of the pp60src kinase was investigated by immunofluorescence microscopy and subcellular fractionation. When permeabilized fibroblasts were stained with a monoclonal anti-34 kd protein antibody, a diffuse reticular pattern was observed. The 34 kd protein was not exposed on the outside surface of the cell. Double immunofluorescence staining experiments established that the 34 kd protein distribution was similar to that of the membrane-associated protein alpha-spectrin. The 34 kd protein was found in cell sections to be concentrated along the cell edges. Taken together, these results suggested that the 34 kd pp60src substrate was associated with the inside surface of the plasma membrane. This conclusion was supported by subcellular fractionation experiments in which the 34 kd protein was observed to fractionate with the plasma membrane. These localization studies support further the hypothesis that many of the primary effects of the pp60src kinase occur at the plasma membrane.
通过免疫荧光显微镜和亚细胞分级分离法研究了pp60src激酶的34kd蛋白底物的亚细胞定位。当用单克隆抗34kd蛋白抗体对通透的成纤维细胞进行染色时，观察到一种弥漫性网状模式。34kd蛋白未暴露在细胞外表面。双重免疫荧光染色实验表明，34kd蛋白的分布与膜相关蛋白α-血影蛋白相似。在细胞切片中发现34kd蛋白集中在细胞边缘。综上所述，这些结果表明34kd pp60src底物与质膜内表面相关。亚细胞分级分离实验支持了这一结论，在该实验中观察到34kd蛋白与质膜分级分离。这些定位研究进一步支持了pp60src激酶的许多主要作用发生在质膜的假说。
