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

TL;DR

This paper discusses a comprehensive approach to mitochondrial proteomics, focusing on preparing mitochondria from mammalian cells, optimizing two-dimensional electrophoresis techniques, and integrating mass spectrometry for protein identification.

Contribution

It introduces specific adjustments to classical 2DE techniques to enhance resolution of mitochondrial proteins in proteomics analysis.

Findings

Improved resolution of mitochondrial proteins using optimized 2DE methods

Effective integration of electrophoresis and mass spectrometry for protein identification

Detailed protocol for mitochondrial protein separation from cultured cells

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.