摘要
目的对小脑及额叶进行比较蛋白质组学研究以探讨其对变性损伤敏感性不同的原因。方法死亡24h内新鲜取材尸检小脑及额叶组织,经提取蛋白质,以固相pH梯度等电聚焦为第一向,SDS PAGE垂直电泳为第二向进行双向电泳(2DE),图像分析软件分析电泳图谱,MALDI TOF质谱或MALDI TOF/TOF串联质谱鉴定蛋白质。结果发现3个蛋白质点在小脑表达量增加,7个蛋白质点在额叶表达量增加。以质谱鉴定了3个小脑中表达量增加的蛋白质点分别为抗氧化蛋白2、肌酸激酶前体(线粒体)和果糖二磷酸醛缩酶C,2个在额叶增加的蛋白质点为胶质纤维酸性蛋白、丙酮酸激酶。30个表达量在额叶及小脑没有显著变化的蛋白质点通过MALDI TOFMS/TOFTOF MSMS和蛋白质数据库检索得到鉴定。结论本研究结果进一步丰富了人脑蛋白质组数据库。额叶与小脑的差异表达蛋白质为深入理解脑老化及变性痴呆分子机制提供了有益的线索。
Objective To compare the results of proteomics between the cerebellum and frontal lobe of the aged. Method Proteins were isolated from human cerebellums and frontal lobes and separated by two-dimensional (2D)gel electrophoresis. The proteins were then stained with silver or colloidal coomassie blue to produce a high-resolution map of the proteome. Selected proteins from this map were in-gel digested with trypsin and the resulting tryptic peptides were analyzed by MALDI-TOF mass spectrometry and MALDI TOF/TOF tandem mass spectrometry. The mass spectrometric data were used to identify the proteins through searches of the SWISSPROT protein sequence database. Results Three up-regulated protein spots in 2D gels of cerebellums were found as compared with those of frontal lobes. These protein spots were identified as antioxidant protein 2, creatine kinase precursor, fructose-bisphosphate aldolase C. Two down-regulated proteins in cerebellums were identified as pyruvate kinase M1 isozyme and glial fibrillary acidic protein. 30 common proteins, the expressions of which were not altered between cerebellums and frontal lobes, were also identified. Conclusions These data will be used for future studies of differential protein expression in neurological disorders. Some proteins are useful for discovering the molecular mechanisms of degenerative dementia and brain aging.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2005年第4期254-257,共4页
Chinese Journal of Internal Medicine
基金
全军"十五"重点课题资助项目(01Z037)
