Proteomic analysis of the wing imaginal discs of Drosophila melanogaster

被引:14
|
作者
Alonso, J [1 ]
Santarén, JF [1 ]
机构
[1] Univ Autonoma Madrid, Fac Ciencias, Ctr Biol Mol Severo Ochoa, CSIC, E-28049 Madrid, Spain
关键词
Drosophila; matrix-assisted laser desorption; ionization-time of flight; two-dimensional gels;
D O I
10.1002/pmic.200400923
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We have combined high-resolution two-dimensional (2-D) gel electrophoresis and mass spectrometry with the aim of identifying proteins represented in the 2-D gel database of the wing imaginal discs of Drosophila melanogaster. First, we obtained a high-resolution 2-D gel pattern of [S-35]methionine + [S-35]cysteine-labeled polypeptides of Schneider cells, a permanent cell line of Drosophila embryonic origin, and compared it with the standard pattern of polypeptides of the wing imaginal disc. These studies reveal qualitative and quantitative differences between the two samples, but have more than 600 polypeptides in common. Second, we carried out preparative 2-D polyacrylamide gel electrophoresis using Schneider cells mixed with radioactively labeled wing imaginal discs in order to isolate some of the shared polypeptides and characterize them by matrix-assisted laser desorption/ionization-time of flight MALDI-TOF analysis. Using this strategy we identified 100 shared proteins represented in the database, and in each case confirmed their identity by MALDI-TOF/TOF analysis.
引用
收藏
页码:474 / 489
页数:16
相关论文
共 50 条