Analysis of Fos protein complexes and Fos-related antigens by high-resolution two-dimensional gel electrophoresis.
Analysis of Fos protein complexes and Fos-related antigens by high-resolution two-dimensional gel electrophoresis.
复制标题
通过高分辨率二维凝胶电泳分析 Fos 蛋白复合物和 Fos 相关抗原。
作者：
B. Franza;L. Sambucetti;D. Cohen;T. Curran
从血清刺激的成纤维细胞和神经生长因子加苯二氮卓类药物处理的嗜铬细胞瘤（PC 12）细胞中分离出含有c-fos蛋白（Fos）和（Fos）相关抗原的蛋白复合物，并通过高分辨率二维凝胶电泳进行研究。结果表明，Fos在成纤维细胞中与碱性39-kDa蛋白（p39）复合，在PC 12细胞中主要与酸性40-kDa蛋白（p40）复合。来自两种细胞类型的全细胞裂解物含有p40，表明Fos和其他细胞蛋白的相互作用取决于细胞的分化状态。除p39和p40外，从任一细胞类型的非变性提取物中分离的Fos复合物中存在约48 kDa的多肽的异质群体。这些蛋白质可能代表一小类Fos结合蛋白。使用抗Fos肽（氨基酸127-152）的抗血清在变性条件下制备的提取物的分析揭示了一系列Fos相关抗原。这些抗原在成纤维细胞和PC 12细胞中被诱导，其中一些具有比Fos更慢的动力学。因此，c-fos可能代表一个基因家族的标记，其中一些是抗原相关的，是早期细胞对不同细胞外刺激的转录反应的一部分。
Protein complexes containing the c-fos protein (Fos) and (Fos)-related antigens were isolated from serum-stimulated fibroblasts and from nerve growth factor plus benzodiazepine-treated pheochromocytoma (PC12) cells, and investigated by high-resolution two-dimensional gel electrophoresis. The results show that Fos is complexed with a basic 39-kDa protein (p39) in fibroblasts, and primarily with an acidic 40-kDa protein (p40) in PC12 cells. Whole cells lysates from both cell types contain p40, suggesting that the interaction of Fos and other cellular proteins is dependent on the differentiated state of the cell. In addition to p39 and p40, a heterogeneous population of polypeptides of approximately 48 kDa are present in Fos complexes isolated from non-denatured extracts of either cell type. These proteins may represent a minor class of Fos-binding proteins. Analysis of extracts prepared under denaturing conditions using antisera raised against a Fos peptide (amino acids 127-152) reveals a series of Fos-related antigens. These antigens are induced, some with a slower kinetics than Fos, in fibroblasts and PC12 cells. Thus, c-fos may represent a marker for a family of genes, some of which are antigenically related, that are part of an early cellular transcriptional response to diverse extracellular stimuli.