Alonso Jana, Santarén Juan F
Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Universidad Autónoma, Cantoblanco, 28049-Madrid, Spain.
J Proteome Res. 2006 Aug;5(8):2025-32. doi: 10.1021/pr0601483.
We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry to identifying proteins represented in a 2-D gel database of Drosophila melanogaster ribosomes. First, we purified ribosomes from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 58 Coomassie blue stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF. Using this strategy we identified 52 ribosomal spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF. The database can be used to analyze Minute mutants of Drosophila.
我们将高分辨率二维(2-D)凝胶电泳与质谱联用,以鉴定果蝇核糖体二维凝胶数据库中所呈现的蛋白质。首先,我们从三龄果蝇幼虫中纯化核糖体,并构建了一个包含58种考马斯亮蓝染色多肽的高分辨率二维凝胶数据库。接下来,我们进行制备性二维聚丙烯酰胺凝胶电泳以分离部分多肽,并通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)对其进行表征。利用这一策略,我们在数据库中鉴定出52个核糖体斑点,并且在每种情况下都通过串联基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/TOF)确认了它们的身份。该数据库可用于分析果蝇的微小突变体。