Abstract
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.
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References
Daum, G., Böhni, P. C., and Schatz, G. (1982) Import of proteins into mitochondria. cytochrome b 2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J. Biol. Chem. 257, 13028–13033.
Hartl, F. U., Ostermann, J., Guiard, B., and Neupert, W. (1987) Successive translocation into and out of the mitochondrial matrix: targeting of proteins to the intermembrane space by a bipartite signal peptide. Cell 51, 1027–1037.
Wiedmann, N., Pfanner, N., and Rahling, P. (2006) Import of precursor proteins into yeast mitochondria. Meth. Mol. Biol. 313, 373–383.
Model, K., Prinz, T., Ruiz, T., et al. (2002) Protein translocase of the outer mitochondrial membrane: role of import receptors in the structural organization of the TOM complex. J. Mol. Biol. 316, 657–666.
Meisinger, C., Sommer, T., and Pfanner, N. (2000) Purification of Saccharomyces cerevisiae mitochondria devoid of microsomal and cytosolic contaminations. Anal. Biochem. 287, 339–342.
Glick, B. S. and Pon, L. A. (1995) Isolation of highly purified mitochondria from Saccharomyces cerevisiae. Methods Enzymol. 260, 213–223.
Martin, H., Eckerskorn, C., Gartner, F., Rassow, J., Lottspeich, F., and Pfanner, N. (1998) The yeast mitochondrial intermembrane space: purification and analysis of two distinct fractions. Anal. Biochem. 265, 123–128.
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.
Sickmann, A., Reinders, J., Wagner, Y., et al. (2003) The proteome of Saccharomyces cerevisiae mitochondria. Proc. Natl. Acad. Sci. USA 100, 13207–13212.
Sikorski, R. S. and Hieter, P. (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27.
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© 2006 Humana Press Inc., Totowa, NJ
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Meisinger, C., Pfanner, N., Truscott, K.N. (2006). Isolation of Yeast Mitochondria. In: Xiao, W. (eds) Yeast Protocol. Methods in Molecular Biology, vol 313. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-958-3:033
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DOI: https://doi.org/10.1385/1-59259-958-3:033
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-58829-437-1
Online ISBN: 978-1-59259-958-5
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