Koseki N, Yoshizato K
Department of Biological Science, Faculty of Science, Hiroshima University, Japan.
Cell Adhes Commun. 1994 Jan;1(4):355-66. doi: 10.3109/15419069409097266.
Cells were cultured on plastic, collagen fibrils or gelatin. General protein synthetic activity of cells did not show any significant difference among the three substrates, whereas the pattern of protein synthesis was substrate-dependent. Profiles of protein synthesis (polypeptide maps) were obtained by subjecting two-dimensional autoradiograms of polyacrylamide gel electrophoresis to a computer-assisted image analyzer. Major polypeptide spots expressed on gelatin were rather like those on plastic. Collagen fibrils caused significant changes in the polypeptides map. Fibroblasts on collagen fibrils produced 364 spots of polypeptides, 26% of which were synthesized specifically on collagen fibrils. The remaining was shared by cells on plastic and was categorized into three groups: (1) polypeptides whose synthesis was up-regulated by collagen fibrils (26% of the total); (2) polypeptides that were expressed equally on both plastic and collagen fibrils (51%); and (3) polypeptides down-regulated by collagen fibrils (23%). A protein with molecular weight of 150 K and an isoelectric point (pI) of 7.3 was one of the collagen-induced and worthy of further analysis. This protein was found to change its pI depending upon the amounts of collagen fibrils and was shown to be located in the mitochondrial fraction.
细胞在塑料、胶原纤维或明胶上培养。细胞的一般蛋白质合成活性在这三种底物之间未显示出任何显著差异，而蛋白质合成模式则依赖于底物。通过对聚丙烯酰胺凝胶电泳的二维放射自显影片进行计算机辅助图像分析，获得蛋白质合成图谱（多肽图谱）。在明胶上表达的主要多肽斑点与在塑料上的相当相似。胶原纤维导致多肽图谱发生显著变化。胶原纤维上的成纤维细胞产生了364个多肽斑点，其中26%是在胶原纤维上特异性合成的。其余的由塑料上的细胞共享，并分为三组：（1）其合成被胶原纤维上调的多肽（占总数的26%）；（2）在塑料和胶原纤维上均同等表达的多肽（51%）；（3）被胶原纤维下调的多肽（23%）。一种分子量为150K、等电点（pI）为7.3的蛋白质是胶原诱导的蛋白质之一，值得进一步分析。发现这种蛋白质的pI会根据胶原纤维的量而变化，并且显示位于线粒体部分。
