小鼠小脑前叶后叶和旁绒球叶的蛋白组学比较分析

PROTEOMICS ANALYSIS OF THE ANTERIOR LOBE,THE POSTERIOR LOBE AND THE PARAFLOCCULUS IN MOUSE CEREBELLUM

目的研究小脑不同区域所含蛋白质表达的差异。方法提取小脑前叶、后叶和旁绒球叶的蛋白质,通过二维凝胶电泳分离蛋白,用PDQuest图像分析软件寻找电泳图谱中的差异蛋白,MALDI TOF/TOF串联质谱分析蛋白质,所得的质谱图通过GPS软件在Mascot数据库中检索并鉴定蛋白质。结果在小脑3个不同区域鉴定了5种差异蛋白,丙酮酸激酶同工酶M2在小脑前叶和后叶的表达量比旁绒球叶的高;核不均一核糖核蛋白L和真核翻译延伸因子2在小脑后叶的表达量少,在小脑前叶和旁绒球叶未检测到;表达序列蛋白AI429145在小脑后叶的表达量高,但在小脑前叶和旁绒球叶未检测到;三磷酸腺苷酶氢离子转运体V0亚单位D异构体1在旁绒球叶的表达量比小脑前叶和后叶的高。结论这些蛋白质在小脑不同区域表达量上的差异可能有助于理解小脑不同区域的特殊功能和疾病。

Objective To study the different expressions of proteins in different regions of cerebellum. Methods Proteins isolated from the anterior lobe, the posterior lobe and the parallocculus of cerebellum were separated by two-dimensional （2-D） gel electrophoresis. Using PDQuest software, proteins on the 2-D maps were selected and further analyzed by matrixassisted laser desorption/ionization two-stage time of flight （MALDI TOF/TOF） tandem mass spectrometry. The mass spectrometric data were used to identify the proteins through the general proteomics standard （GPS） search and Mascot database. Results There were five proteins identified in three regions of cerebella. The level of pyruvate kinase isozyme M2 was higher in the anterior and posterior lobes of cerebellum than in the paraflocculus. Heterogeneous nuclear ribonucleoprotein L and eukaryotic translation elongation factor 2 were only identified in the posterior lobe with low expression, and were not detected in other regions. The expressed sequence AI429145 was only detected in the posterior lobe. The paraflocculus expressed higher level of ATPase （H^＋ transporting, VO subunit D, isoform 1 ） compared with the anterior and posterior lobes. Conclusion The different expressions of those proteins may be important for the understanding of the specific functions and diseases in different regions of cerebellum.