Isolation of Yeast Mitochondria
Often preparations of isolated organelles contain other, unwanted, cellular components. For biochemical experiments to determine the localization of newly identified proteins, or to determine the whole set of proteins (or the proteome) from a desired organelle, these unwanted components often confuse the resulting data. For these types of studies, it is crucial to have highly pure fractions of the desired organelle. Here we describe a protocol for purification of mitochondria from Saccharomyces cerevisiae cells devoid of contamination from other cellular compartments.
KeywordsSucrose Gradient Homogenization Buffer Yeast Mitochondrion Resuspend Pellet Mitochondrial Sample
- 3.Wiedmann, N., Pfanner, N., and Rahling, P. (2006) Import of precursor proteins into yeast mitochondria. Meth. Mol. Biol. 313, 373–383.Google Scholar
- 8.Meisinger, C., Ryan, M. T., Hill, K., et al. (2001) The protein import channel of the outer mitochondrial membrane: a highly stable Tom40-Tom22 core structure differentially interacts with preproteins, small Tom proteins and import receptors. Mol. Cell. Biol. 21, 2337–2348.PubMedCrossRefGoogle Scholar