Celis J E, Crüger D, Kiil J, Dejgaard K, Lauridsen J B, Ratz G P, Basse B, Celis A, Rasmussen H H, Bauw G
Institute of Medical Biochemistry, Aarhus University, Denmark.
Electrophoresis. 1990 Mar;11(3):242-54. doi: 10.1002/elps.1150110308.
A two-dimensional (2-D) gel database of proteins from noncultured total normal human epidermal keratinocytes has been established. A total of 1449 [35S]methionine labelled proteins (1112 isoelectric focusing, 337 nonequilibrium pH gradient electrophoresis) were resolved and recorded using computer assisted (PDQ-SCAN and PDQUEST software) 2-D gel electrophoresis. By matching the protein patterns of total keratinocytes and transformed human amnion cells (master database; Celis et al., Leukemia 1988, 2, 561-602) as well as by 2-D immunoblotting and microsequencing of keratinocyte proteins, it was possible to identify 72 polypeptides in the keratinocyte database. The database also includes data on polypeptides that are synthesized at a higher level by keratinocytes enriched in basal cells, and on six secreted proteins which are produced, albeit at a reduced rate, by normal keratinocytes and that are strongly up-regulated in psoriatic epidermis (Celis et al., FEBS Letters, in press).
已建立了来自未培养的正常人表皮角质形成细胞蛋白质的二维(2-D)凝胶数据库。使用计算机辅助(PDQ-SCAN和PDQUEST软件)二维凝胶电泳分离并记录了总共1449种[35S]甲硫氨酸标记的蛋白质(1112种等电聚焦,337种非平衡pH梯度电泳)。通过匹配总角质形成细胞和转化的人羊膜细胞的蛋白质模式(主数据库;Celis等人,《白血病》1988年,2卷,561 - 602页),以及通过角质形成细胞蛋白质的二维免疫印迹和微量测序,有可能在角质形成细胞数据库中鉴定出72种多肽。该数据库还包括关于在富含基底细胞的角质形成细胞中高水平合成的多肽的数据,以及关于六种分泌蛋白的数据,这些蛋白由正常角质形成细胞产生,尽管速率降低,并且在银屑病表皮中强烈上调(Celis等人,《欧洲生物化学学会联合会快报》,即将发表)。