Characterization of the Drosophila melanogaster mitochondrial proteome

J Proteome Res. Sep-Oct 2005;4(5):1636-45. doi: 10.1021/pr050130c.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / metabolism*
  • Electrophoresis, Gel, Two-Dimensional
  • Hydrogen-Ion Concentration
  • Image Processing, Computer-Assisted
  • Immunoblotting
  • Isoelectric Focusing
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Mitochondria / metabolism
  • Mitochondrial Proteins / chemistry
  • Molecular Sequence Data
  • Peptides / chemistry
  • Proteome*
  • Proteomics / methods*
  • Software
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Mitochondrial Proteins
  • Peptides
  • Proteome