摘要
使用双向电泳 (2 DE)比较成年和老年小鼠脑蛋白质差异 ,从分子水平初步探索老年脑蛋白整体变化规律 .以固相 pH梯度等电聚焦为第一向 ,SDS 聚丙烯酰胺凝胶水平电泳 (PAGE)为第二向进行 2 DE .图象分析软件Imagemaster 2DElite分析电泳图谱 .重复性实验结果显示 ,同组样品在三次不同实验中所得蛋白质斑点数目的相对标准差 (变异系数 )为 4 43%± 0 2 5 % ;同一蛋白质斑点在三次实验中等电点、分子质量和蛋白质量的相对标准差分别为 8 76 %± 5 14% ,13 0 0 %± 4 2 2 %和 10 84%± 9 16 % .成年和老年小鼠脑组织 2 DE图谱分别获得 996和 12 5 6个蛋白质斑点 ,其中 8个蛋白质在老年脑组织中含量降低 ,2 0个蛋白质斑点含量增加 .另至少有 4个蛋白质斑点在老年脑组织中缺失 ,14个蛋白质点为老年脑特有 .
A wide variety alterations are common in brains of aged individuals. Proteomatic analysis is anew power toot to reveal such changes at protein level. The two-dimensional electrophoresis (2-DE) was employed to compare the global protein patterns between adult and aged murine brains. One hundred and fifty micrograms of murine brain protein extracted with 5 mol/L urea, 2 mol/L thiourea, 2% CHAPS plus 2% SB3-10 was run immobilized pH gradient (IPG) isoelectric focusing electrophoresis as the first dimension, and then horizontal SDS-PAGE as the second dimension. The relative standard deviations (coefficient of variation) for protein number, pI, molecular mass and volume of protein in three different experiments were 4.43% +/- 0.25%, 8.76% +/- 5.14%, 13.00% +- 4.22% and 10.84% +/- 9.16%, respectively. Totally, 996 and 1 256 protein spots were obtained in adult and aged murine brain map, respectively, of which 8 spots increased and 20 spots decreased in quantity. Furthermore, 4 spots were missing and 14 new spots emerged in aged murine brain compared with adult mice. The differentially displayed proteins between adult and aged murine brain are useful for diagnosing brain degenerative disorders.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2001年第5期683-687,共5页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助 (3 992 80 15 )~~
