title: Characterization of the Drosophila melanogaster mitochondrial proteome.
abstract: 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.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049-Madrid, Spain.
authors.forename: Jana
authors.initials: J
authors.lastname: Alonso
authors.forename: Javier M
authors.initials: JM
authors.lastname: Rodriguez
authors.forename: Luis Alberto
authors.initials: LA
authors.lastname: Baena-López
authors.forename: Juan F
authors.initials: JF
authors.lastname: Santarén
mesh_terms: 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
chemical_list: D024101:Mitochondrial Proteins; D010455:Peptides; D020543:Proteome