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Methods Cell Biol. 2013;117:411-31. doi: 10.1016/B978-0-12-408143-7.00022-0.

Identification of multimolecular complexes and supercomplexes in compartment-selective membrane microdomains.

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  • 1Department of Microbiology and Immunology, McGill University, Montreal, QC, Canada.

Abstract

Cellular membranes contain specialized microdomains that play important roles in a wide range of cellular processes. These microdomains can be found in the plasma membrane and other membranes within the cell. Initially labeled lipid rafts and defined as being resistant to extraction by nonionic detergents and enriched in cholesterol and glycosphingolipids, we now understand that these membrane microdomains are very dynamic and heterogeneous membrane structures whose composition and function can vary widely depending on their cellular location. Indeed, though they are classically associated with the plasma membrane and have been shown to facilitate a wide variety of processes, including signal transduction and membrane trafficking, specialized membrane microdomains have also been identified in other membranes including those in the mitochondria. These mitochondrial membrane microdomains are enriched in cardiolipin, the signature phospholipid of the mitochondria, and may have important implications in metabolism by facilitating optimal assembly and function of the mitochondrial respiratory chain. Furthermore, isolation of multimolecular complexes while retaining their supramolecular interactions has been critical to the study of mitochondrial respiratory supercomplexes. Here, we discuss methods to isolate various membrane microdomains, including detergent-insoluble glycosphingolipid microdomains, mitochondrial cardiolipin-enriched microdomains, and blue-native gel electrophoresis of mitochondrial membranes.

Copyright © 2013 Elsevier Inc. All rights reserved.

KEYWORDS:

Blue-native gel electrophoresis; Cardiolipin; Detergent-insoluble microdomains; Glycosphingolipids; Mitochondria; Mitochondrial membranes; Mitochondrial supercomplexes; Plasma membrane

PMID:
24143990
[PubMed - indexed for MEDLINE]
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