Bravo R, Celis J E
Clin Chem. 1982 Apr;28(4 Pt 2):949-54.
We resolved the [35S]methionine polypeptides synthesized by normal [lung fibroblasts (WI-38), amnion cells] and transformed [SV40 transformed WI-38, AMA (spontaneously transformed amnion cells)] cultured human cells, using two-dimensional gel electrophoresis under conditions in which about 1300 polypeptides could be reproducibly separated. These studies demonstrated important changes in the relative proportions of several polypeptides that are present both in normal and transformed cells. Of a total of 400 common polypeptides that we quantitated for each cell type, 53 (22 basic and 31 acidic) varied by 40% of more in both cell pairs. Among these, we have identified vimentin (IEF 26), cyclin (IEF 49), and a tropomyosin-related polypeptide (IEF 52). No new major polypeptide was detected in the transformed cells, at least at the level of resolution currently achieved by this technique. Similar but qualitative studies of [32P]orthophosphate-labeled proteins revealed that, of 250 analyzed, only seven common phosphoproteins, including phosphovimentin (IEF 26e), varied consistently in both cell pairs. These results strengthen our previous conclusion that transformation results in changes in the relative proportions of polypeptides synthesized in normal and transformed cells rather than in the appearance of new polypeptides in transformed cells.
我们使用二维凝胶电泳，在可重复分离约1300种多肽的条件下，解析了正常[肺成纤维细胞（WI - 38）、羊膜细胞]和转化的[SV40转化的WI - 38、AMA（自发转化的羊膜细胞）]培养的人细胞合成的[35S]甲硫氨酸多肽。这些研究表明，正常细胞和转化细胞中都存在的几种多肽的相对比例发生了重要变化。在我们为每种细胞类型定量的总共400种常见多肽中，53种（22种碱性和31种酸性）在两组细胞中的变化都超过了40%。其中，我们鉴定出波形蛋白（IEF 26）、细胞周期蛋白（IEF 49）和一种原肌球蛋白相关多肽（IEF 52）。至少在目前该技术所达到的分辨率水平上，在转化细胞中未检测到新的主要多肽。对[32P]正磷酸盐标记蛋白的类似但定性的研究表明，在分析的250种蛋白中，只有7种常见的磷蛋白，包括磷酸波形蛋白（IEF 26e），在两组细胞中都有一致的变化。这些结果强化了我们之前的结论，即转化导致正常细胞和转化细胞中合成的多肽相对比例发生变化，而不是在转化细胞中出现新的多肽。
