Proteomic analysis of the wing imaginal discs of Drosophila melanogaster
Proteomic analysis of the wing imaginal discs of Drosophila melanogaster
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DOI：
10.1002/pmic.200400923
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发表时间：
2005-02-01
期刊：
影响因子：
3.4
通讯作者：
Santarén, JF
中科院分区：
文献类型：
--
作者：
Alonso, J;Santarén, JF
我们结合了高分辨率的二维（2-D）凝胶电泳和质谱，目的是识别蛋白质的2-D凝胶数据库中的果蝇翅成虫盘。首先，我们获得了[S-35]甲硫氨酸+ [S-35]半胱氨酸标记的Schneider细胞（果蝇胚胎起源的永久细胞系）多肽的高分辨率2-D凝胶图案，并将其与翼成虫盘多肽的标准图案进行了比较。这些研究揭示了两种样品之间的定性和定量差异，但有超过600种多肽是共同的。第二，我们进行了制备2-D聚丙烯酰胺凝胶电泳使用施耐德细胞混合放射性标记的翼成虫盘，以分离出一些共享的多肽，并通过基质辅助激光解吸/电离飞行时间MALDI-TOF分析表征它们。使用这种策略，我们确定了100个共享的蛋白在数据库中表示，并在每种情况下，确认其身份的MALDI-TOF/TOF分析。
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.