Detergent-Resistant Membranes and the Use of Cholesterol Depletion

Sebastian Schuck, Masanori Honsho, Kai Simons

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Isolation of detergent-resistant membranes (DRMs) is a simple and useful biochemical method for analyzing the possible raft association of proteins and lipids. Raft proteins do not become solubilized, but remain associated with lipids. As a result, they have a lower apparent density than detergent-soluble membrane proteins and can be isolated by equilibrium density. Detergent solubility of an otherwise detergent-insoluble protein after cholesterol depletion with methyl-~-cyclodextrin can therefore serve as an additional criterion for raft association. However, the composition of DRMs may only imperfectly reflect the association of membrane components with lipid rafts in cell membranes. Cyclodextrin might be more effective on cell homogenate than on living cells, but the reasons for this are not completely clear. Various density gradient media can be used, and the choice between sucrose and iodixanol is largely a question of personal preference. However, sucrose might be superior if lipids are to be analyzed by a very sensitive method such as mass spectrometry, as iodixanol seems to have a weak tendency to follow lipids during extraction from DRMs. © 2006

Original languageEnglish
Title of host publicationCell Biology, Four-Volume Set
PublisherElsevier Inc.
Pages5-9
Number of pages5
Volume2
ISBN (Print)9780121647308
DOIs
Publication statusPublished - Dec 1 2006
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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