Proteomic analysis of the wing imaginal discs of Drosophila melanogaster.
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 [35S]methionine + [35S]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.
Alonso
Jana
J
Centro de Biología Molecular "Severo Ochoa", CSIC-UAM, Universidad Autónoma de Madrid, Madrid, Spain.
Santarén
Juan F
JF
D000818:Animals; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D016208:Databases, Factual; D029721:Drosophila Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D004331:Drosophila melanogaster / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D007091:Image Processing, Computer-Assisted; D007814:Larva; D013058:Mass Spectrometry; D008715:Methionine / Q000378:metabolism; D008954:Models, Biological; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013462:Sulfur Radioisotopes / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014921:Wings, Animal / Q000737:chemistry*
D029721:Drosophila Proteins; D020543:Proteome; D013462:Sulfur Radioisotopes; D008715:Methionine; D014357:Trypsin; D003545:Cysteine