Mitochondrial proteomics : analysis of a whole mitochondrial extract with two-dimensional electrophoresis.
Mitochondria are complex organelles, and their proteomics analysis requires a combination of techniques. The emphasis in this chapter is made first on mitochondria preparation from cultured mammalian cells, then on the separation of the mitochondrial proteins with two-dimensional electrophoresis (2DE), showing some adjustment over the classical techniques to improve resolution of the mitochondrial proteins. This covers both the protein solubilization, the electrophoretic part per se, and the protein detection on the gels, which makes the interface with the protein identification part relying on mass spectrometry.
Rabilloud
Thierry
T
CEA/DSV/IRTSV/BBSI, UMR CNRS 5092, Biologie et Biophysique des systèmes intégrés, CEA Grenoble, 17 rue des martyrs, Grenoble, France.
D000818:Animals; D015153:Blotting, Western; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007425:Intracellular Membranes / Q000737:chemistry / Q000648:ultrastructure; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008930:Mitochondria, Liver / Q000737:chemistry* / Q000648:ultrastructure; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011489:Protein Denaturation; D020543:Proteome
D024101:Mitochondrial Proteins; D020543:Proteome