Protein profiling of rat cerebella during development
Protein profiling of rat cerebella during development
复制标题
大鼠小脑发育过程中的蛋白质分析
作者：
M. Taoka;A. Wakamiya;H. Nakayama;T. Isobe
通过双向凝胶电泳（2-DE）和质谱（MS）分析发育中大鼠小脑的蛋白质谱。对成年大鼠小脑的分析产生了一个蛋白质图谱，该图谱包括pH范围为3 - 10、质量范围为8 - 100 kDa的高分辨率2-DE后银染色可检测到的约3000个斑点。为了获得比较小脑蛋白发育特征的标志，使用基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF-MS）对100个点进行肽质量指纹图谱分析，并在图上分配67个点。发育中的小脑的轮廓分析显示，在发展过程中的蛋白质表达的显着变化。在大多数情况下，蛋白质的表达水平随着小脑的成熟而增加，而42个斑点的表达在未成熟小脑中出现特异性或显着丰富。这些点的肽质量指纹图谱使我们能够识别29种蛋白质，除了功能未知的蛋白质外，还包括许多已知在中枢神经系统发育中起作用的蛋白质。这些结果表明，蛋白质组学的方法是有价值的大规模鉴定参与小脑形态发生的蛋白质。
Protein profiles of developing rat cerebella were analyzed by means of two‐dimensional gel electrophoresis (2‐DE) and mass spectrometry (MS). The analysis of adult rat cerebellum gave rise to a protein map comprising ˜3000 spots detectable by silver staining following high resolution 2‐DE with a pH range of 3‐10 and a mass range of 8‐100 kDa. To obtain landmarks for comparison of developmental profiles of cerebellar proteins, 100 spots were subjected to peptide mass fingerprinting using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS), and 67 spots were assigned on the map. Analysis of profiles of the developing cerebella revealed significant changes in the expression of proteins during development. In most cases the expression levels of proteins increased as the cerebellum matured, while the expression of 42 spots appeared specific or remarkably abundant in the immature cerebellum. Peptide mass fingerprinting of these spots allowed us to identify 29 proteins, which include, in addition to proteins of unknown function, many proteins known to have roles in the development of the central nervous system. These results suggest that the proteomic approach is valuable for mass identification of proteins involved in cerebellar morphogenesis.