Characterization of the Drosophila melanogaster mitochondrial proteome.
We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry with the aim of identifying proteins represented in the 2-D gel database of Drosophila melanogaster mitochondria. First, we purified mitochondria from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 231 silver-stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF analysis. Using this strategy, we identified 66 mitochondrial spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF analysis. In addition, we generated antibodies against two of the mitochondrial proteins as tools for characterizing the organelle.
Alonso
Jana
J
Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049-Madrid, Spain.
Rodriguez
Javier M
JM
Baena-López
Luis Alberto
LA
Santarén
Juan F
JF
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D024101:Mitochondrial Proteins; D010455:Peptides; D020543:Proteome