Because virtually all cellular processes are based on proteins, detailed knowledge of the mitochondrial proteome represents an integral part of understanding mitochondrial function. The analysis of very complex protein mixtures such as entire cell organelles makes high demands on analysis techniques in order to ensure integrity of the obtained data set. The use ofSaccharomyces cerevisiaeas a model system allows the isolation of mitochondria of utmost purity in large amounts. Various approaches can be accomplished in the subsequent analysis to achieve the most complete overview possible. Combinations of orthogonal proteomics techniques include two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D-SDS-PAGE), and nano-LC-MS/MS (nano-liquid chromatography with tandem mass spectrometry) as well as multidimensional high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The inherent limitations of the individual methods can be countervailed by parallel application of these approaches
