Isolation and characterization of highly purified mitochondrial outer membranes of the yeast Saccharomyces cerevisiae (method).

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  • Additional Information
    • Source:
      Publisher: Informa Healthcare Country of Publication: England NLM ID: 9430797 Publication Model: Print Cited Medium: Print ISSN: 0968-7688 (Print) Linking ISSN: 09687688 NLM ISO Abbreviation: Mol Membr Biol Subsets: MEDLINE
    • Publication Information:
      Publication: London : Informa Healthcare
      Original Publication: London, UK : Taylor & Francis, c1994-2019.
    • Subject Terms:
      Intracellular Membranes*/metabolism ; Mitochondria* ; Saccharomyces cerevisiae*; Cell Fractionation/*methods; Organelles ; Phospholipids/metabolism
    • Abstract:
      Mitochondrial outer membrane vesicles (OMV) from the yeast Saccharomyces cerevisiae were prepared by osmotic swelling and mechanical disruption of mitochondria that had been isolated at pH 6.0 and purified by density gradient centrifugation. The OMV were obtained in a yield of 1% (protein/protein) with respect to the mitochondria. The OMV were shown to be essentially free of mitochondrial inner membrane protein markers, while contamination with endoplasmic reticulum was around 5% (protein-based). The very low phosphatidylserine synthase activity present in the OMV preparation indicated that contamination with mitochondria-associated membranes (MAM) was negligible. The resistance of the outer membrane protein Tom40 to digestion by trypsin demonstrated the sealed nature and right-side out orientation of the OMV. Analysis of the phospholipid composition revealed that the contents of phosphatidylcholine and phosphatidylinositol are higher and the content of phosphatidylethanolamine is lower in the mitochondrial outer membrane as compared to whole mitochondria. Cardiolipin is largely depleted in the OMV.
    • Accession Number:
      0 (Phospholipids)
    • Publication Date:
      Date Created: 19990727 Date Completed: 19991014 Latest Revision: 20191103
    • Publication Date:
      20231215
    • Accession Number:
      10.1080/096876899294670
    • Accession Number:
      10417986