Toda T, Kaji K, Kimura N
Department of Molecular Biology, Tokyo Metropolitan Institute of Gerontology, Japan.
Electrophoresis. 1998 Feb;19(2):344-8. doi: 10.1002/elps.1150190232.
Cellular proteins of a normal human diploid fibroblast line (TIG-3) at various stages of replicative aging were resolved by horizontal isoelectric focusing on an immobilized pH gradient, followed by vertical sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). Spot proteins were visualized by silver staining and quantitated by image processing. All corresponding spots were matched among two-dimensional gel images, and variation profiles in relative abundance of individual proteins during in vitro aging were classified into five categories, i.e., (i) increase, (ii) decrease, (iii) increase followed by decrease, (iv) decrease followed by increase, and (v) irregular or nonsignificant variation. The new concept of the Tokyo Metropolitan Institute of Gerontology two-dimensional gel protein database (TMIG-2DPAGE) was prepared from the above data to support research on cellular aging. The database was put on our World Wide Web home page at the URL of http://www.tmig.or.jp/2D/ to allow free access through the Internet. The individual protein data entries were linked to the standard spot protein map of the two-dimensional gel image in order to be accessible by clicking the mouse on it.
采用固定化pH梯度水平等电聚焦，随后进行垂直十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶电泳（PAGE），对正常人二倍体成纤维细胞系（TIG-3）在复制性衰老不同阶段的细胞蛋白质进行分离。斑点蛋白通过银染可视化，并通过图像处理进行定量。在二维凝胶图像之间匹配所有相应的斑点，并将体外衰老过程中单个蛋白质相对丰度的变化谱分为五类，即：（i）增加，（ii）减少，（iii）先增加后减少，（iv）先减少后增加，以及（v）不规则或无显著变化。根据上述数据编制了东京都老人医学研究所二维凝胶蛋白质数据库（TMIG-2DPAGE）的新概念，以支持细胞衰老研究。该数据库发布在我们的万维网主页上，网址为http://www.tmig.or.jp/2D/，可通过互联网免费访问。单个蛋白质数据条目与二维凝胶图像的标准斑点蛋白质图谱相关联，以便通过鼠标点击进行访问。
