Mitochondrial proteomics: analysis of a whole mitochondrial extract with two-dimensional electrophoresis

Abstract

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.

References in corpus (5)

• Evaluation of nonionic and zwitterionic detergents as membrane protein solubilizers in two-dimensional electrophoresis
• Progress in the definition of a reference human mitochondrial proteome
• About the mechanism of interference of silver staining with peptide mass spectrometry
• About thiol derivatization and resolution of basic proteins in two-dimensional electrophoresis
• Alterations of the mitochondrial proteome caused by the absence of mitochondrial DNA: A proteomic view