Pollak Daniela, Krapfenbauer Kurt, Fountoulakis Michael, Peyrl Andreas, Lubec Gert
Department of Pediatrics, Division Basic Science, University of Vienna, Währinger Gürtel 18, A-1090 Vienna, Austria.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Sep 5;808(2):185-208. doi: 10.1016/j.jchromb.2004.05.008.
Although a variety of signaling systems and signaling proteins have been described, cell specific expression of these structures has not yet been systematically studied. Human amnion, bronchial epithelial, fibroblast, glial, kidney, lymphocyte and mesothelial cells were subjected to two-dimensional-gel electrophoresis followed by analysis of protein spots by MALDI-TOF and subsequent identification by specific software. A series of well-documented signaling proteins showed cell specific expressional patterns. Five hypothetical proteins--hypothetical 37.5 kDa protein, similar to calsyntenin 1, hypothetical armadillo repeat/plakoglobulin ARM-repeat profile containing protein, 11 days embryo cDNA clone 2700084k13, hypothetical protein flj22171--so far predicted from their nucleic acid sequence only, were identified, complementing already reported signaling cascades. An analytical tool for the concomitant determination of a large series of signaling structures by an antibody independent protein-chemical method is provided.
尽管已经描述了多种信号系统和信号蛋白,但尚未对这些结构的细胞特异性表达进行系统研究。对人羊膜、支气管上皮、成纤维细胞、神经胶质细胞、肾、淋巴细胞和间皮细胞进行二维凝胶电泳,然后通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)分析蛋白斑点,并随后使用特定软件进行鉴定。一系列有充分文献记载的信号蛋白显示出细胞特异性表达模式。鉴定出了五种目前仅根据其核酸序列预测的假定蛋白——假定的37.5 kDa蛋白,类似于钙联蛋白1、假定的含有犰狳重复/桥粒斑珠蛋白ARM重复谱的蛋白、11天胚胎cDNA克隆2700084k13、假定蛋白flj22171,补充了已报道的信号级联反应。提供了一种通过独立于抗体的蛋白质化学方法同时测定大量信号结构的分析工具。