Isolation of plasma membrane-associated membranes from rat liver.

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  • Additional Information
    • Source:
      Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101284307 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1750-2799 (Electronic) Linking ISSN: 17502799 NLM ISO Abbreviation: Nat Protoc Subsets: MEDLINE
    • Publication Information:
      Original Publication: London, UK : Nature Pub. Group, 2006-
    • Subject Terms:
      Cell Fractionation/*methods ; Cell Membrane/*physiology ; Endoplasmic Reticulum/*physiology ; Histocytological Preparation Techniques/*methods ; Liver/*cytology; Animals ; Centrifugation/methods ; Rats
    • Abstract:
      Dynamic interplay between intracellular organelles requires a particular functional apposition of membrane structures. The organelles involved come into close contact, but do not fuse, thereby giving rise to notable microdomains; these microdomains allow rapid communication between the organelles. Plasma membrane-associated membranes (PAMs), which are microdomains of the plasma membrane (PM) interacting with the endoplasmic reticulum (ER) and mitochondria, are dynamic structures that mediate transport of proteins, lipids, ions and metabolites. These structures have gained much interest lately owing to their roles in many crucial cellular processes. Here we provide an optimized protocol for the isolation of PAM, PM and ER fractions from rat liver that is based on a series of differential centrifugations, followed by the fractionation of crude PM on a discontinuous sucrose gradient. The procedure requires ∼8-10 h, and it can be easily modified and adapted to other tissues and cell types.
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    • Grant Information:
      GGP11139 Italy TI_ Telethon
    • Publication Date:
      Date Created: 20140118 Date Completed: 20140915 Latest Revision: 20220129
    • Publication Date:
      20231215
    • Accession Number:
      10.1038/nprot.2014.016
    • Accession Number:
      24434800