Moore H P, Kelly R B
J Cell Biol. 1985 Nov;101(5 Pt 1):1773-81. doi: 10.1083/jcb.101.5.1773.
The mouse pituitary cell line, AtT-20, packages the adrenocorticotropic hormone (ACTH) in secretory vesicles and releases it when the cell is stimulated with secretagogues. These cells have the capacity, after transfection with the appropriate DNA, to package heterologous peptide hormones into the regulated secretory vesicles (Moore, H. P. H., M. D. Walker, F. Lee, and R. B. Kelly, 1983, Cell, 35:531-538). To test if other secreted proteins prefer a different route to the surface, we have transfected AtT-20 cells with DNAs coding for a fragment of a membrane protein, the vesicular stomatitis virus G protein from which the membrane spanning domain has been deleted (Rose, J. K., and J. E. Bergmann, 1982, Cell, 17:813-819). We found that the secreted vesicular stomatitis virus G proteins were not transported to the regulated secretory vesicles. Instead they preferentially exited the cell by the constitutive pathway previously found in these cells (Gumbiner, B., and R. B. Kelly, 1982, Cell, 28:51-59). In contrast, human growth hormone transfected into the cells by the same procedure was transported to the regulated pathway with a similar efficiency as the endogenous hormone ACTH. Transport of the secreted G protein to the regulated pathway, if it occurs at all, is at least 30-fold less efficient than peptide hormones. We conclude that the transport machinery in AtT-20 cells must selectively recognize different secreted proteins and sort them into distinct secretory pathways.
小鼠垂体细胞系AtT - 20将促肾上腺皮质激素(ACTH)包装在分泌囊泡中,并在细胞受到促分泌剂刺激时释放它。在用适当的DNA转染后,这些细胞有能力将异源肽激素包装到调节性分泌囊泡中(Moore, H. P. H., M. D. Walker, F. Lee, and R. B. Kelly, 1983, Cell, 35:531 - 538)。为了测试其他分泌蛋白是否偏好不同的表面运输途径,我们用编码膜蛋白片段的DNA转染了AtT - 20细胞,该膜蛋白是已缺失跨膜结构域的水泡性口炎病毒G蛋白(Rose, J. K., and J. E. Bergmann, 1982, Cell, 17:813 - 819)。我们发现分泌的水泡性口炎病毒G蛋白没有被运输到调节性分泌囊泡中。相反,它们优先通过这些细胞中先前发现的组成型途径离开细胞(Gumbiner, B., and R. B. Kelly, 1982, Cell, 28:51 - 59)。相比之下,通过相同程序转染到细胞中的人生长激素以与内源性激素ACTH相似的效率被运输到调节性途径。分泌的G蛋白运输到调节性途径(如果确实发生的话)的效率至少比肽激素低30倍。我们得出结论,AtT - 20细胞中的运输机制必须选择性地识别不同的分泌蛋白,并将它们分类到不同的分泌途径中。