Isolation of plasma membrane-associated membranes from rat liver
- PMID: 24434800
- DOI: 10.1038/nprot.2014.016
Isolation of plasma membrane-associated membranes from rat liver
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.
Similar articles
-
Protocol for the isolation and purification of endoplasmic reticulum-plasma membrane junctions from the mouse brain.STAR Protoc. 2024 Sep 20;5(3):103253. doi: 10.1016/j.xpro.2024.103253. Epub 2024 Aug 10. STAR Protoc. 2024. PMID: 39126654 Free PMC article.
-
Isolation of mitochondria-associated membranes and mitochondria from animal tissues and cells.Nat Protoc. 2009;4(11):1582-90. doi: 10.1038/nprot.2009.151. Epub 2009 Oct 8. Nat Protoc. 2009. PMID: 19816421
-
Isolation of subcellular fractions from the yeast Saccharomyces cerevisiae.Curr Protoc Cell Biol. 2001 May;Chapter 3:Unit 3.8. doi: 10.1002/0471143030.cb0308s08. Curr Protoc Cell Biol. 2001. PMID: 18228360
-
Endoplasmic reticulum-plasma membrane contacts: Principals of phosphoinositide and calcium signaling.Curr Opin Cell Biol. 2020 Apr;63:125-134. doi: 10.1016/j.ceb.2020.01.010. Epub 2020 Feb 20. Curr Opin Cell Biol. 2020. PMID: 32088611 Review.
-
Endoplasmic reticulum-plasma membrane junctions: structure, function and dynamics.J Physiol. 2016 Jun 1;594(11):2837-47. doi: 10.1113/JP271142. Epub 2016 May 7. J Physiol. 2016. PMID: 26939537 Free PMC article. Review.
Cited by
-
Dynamic Remodeling of Membranes and Their Lipids during Acute Hormone-Induced Steroidogenesis in MA-10 Mouse Leydig Tumor Cells.Int J Mol Sci. 2021 Mar 4;22(5):2554. doi: 10.3390/ijms22052554. Int J Mol Sci. 2021. PMID: 33806352 Free PMC article.
-
Spectrin-beta 2 facilitates the selective accumulation of GABAA receptors at somatodendritic synapses.Commun Biol. 2023 Jan 5;6(1):11. doi: 10.1038/s42003-022-04381-x. Commun Biol. 2023. PMID: 36604600 Free PMC article.
-
Biomimetic Nanoparticles Camouflaged in Cancer Cell Membranes and Their Applications in Cancer Theranostics.Front Oncol. 2020 Jan 21;9:1560. doi: 10.3389/fonc.2019.01560. eCollection 2019. Front Oncol. 2020. PMID: 32039028 Free PMC article.
-
A caveolin-1 dependent glucose-6-phosphatase trafficking contributes to hepatic glucose production.Mol Metab. 2023 Apr;70:101700. doi: 10.1016/j.molmet.2023.101700. Epub 2023 Mar 2. Mol Metab. 2023. PMID: 36870604 Free PMC article.
-
LDL Promotes Disorders in β-Cell Cholesterol Metabolism, Implications on Insulin Cellular Communication Mediated by EVs.Metabolites. 2022 Aug 16;12(8):754. doi: 10.3390/metabo12080754. Metabolites. 2022. PMID: 36005626 Free PMC article.
References
Publication types
MeSH terms
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous