Institute of Hygiene and Occupational Medicine, University Hospital Essen, Essen, Germany.
Electrophoresis. 2011 Dec;32(24):3600-11. doi: 10.1002/elps.201100220.
Epithelial tissue lining the inner side of the urinary bladder is the most common target for bladder cancer-related diseases. Bladders of freshly slaughtered pigs were utilised for a comprehensive analysis of the proteome and phosphoproteome of bladder epithelial cells. Following protein separation by 2-D gel electrophoresis and identification by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) the first proteome and phosphoproteome maps of pig urinary bladder epithelial cells (PUBEC) were established. A total of 120 selected protein spots were identified. By using the La(3+) enrichment method further developed in our laboratory we identified 31 phosphoproteins with minimal contamination by non-phosphopeptides. The 2-DE map of pig urothelial cells may prove as a useful tool for studies on uroepithelial biology, and the analysed phosphoproteins expression pattern, together with the whole cell proteome, will be helpful for identifying the proteins involved in bladder-related diseases.
衬在膀胱内侧的上皮组织是膀胱癌相关疾病最常见的靶标。利用刚宰杀的猪的膀胱对膀胱上皮细胞的蛋白质组和磷酸化蛋白质组进行了全面分析。通过二维凝胶电泳分离蛋白质,并用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)进行鉴定,建立了猪膀胱上皮细胞(PUBEC)的第一张蛋白质组和磷酸化蛋白质组图谱。共鉴定出 120 个选定的蛋白质斑点。通过使用我们实验室进一步开发的 La(3+)富集方法,我们鉴定出了 31 种磷酸化蛋白,其中非磷酸肽的污染最小。猪尿路上皮细胞的 2-DE 图谱可能成为研究尿路上皮生物学的有用工具,而分析的磷酸化蛋白表达模式,以及整个细胞蛋白质组,将有助于鉴定与膀胱相关疾病相关的蛋白质。