Intracellular localization of organized lipid domains of C16-ceramide/cholesterol

Michal Goldschmidt-Arzi, Eyal Shimoni, Helena Sabanay, Anthony H. Futerman, Lia Addadi

Research output: Contribution to journalArticlepeer-review

Abstract

Lipid microdomains, also called lipid rafts, consisting of sphingolipids and cholesterol, play important roles in membrane trafficking and in signaling. Despite years of study of the composition, size, half-life and dynamic organization of these domains, many open questions remain about their precise characteristics. To address some of these issues, we have developed a new experimental approach involving the use of specific monoclonal antibodies as recognition tools. One such antibody was raised against a homogeneous, mixed, ordered monolayer phase comprised of 60:40 mol% cholesterol:C16-ceramide, and has been used previously to demonstrate the existence of C16-ceramide/cholesterol domains in the membranes of cultured cells. We now use a combination of quantitative fluorescence microscopy, immuno-transmission electron microscopy and immuno-scanning cryo-electron microscopy, optimized for the study of intracellular lipid antigens. In a variety of cultured cells, C16-ceramide/cholesterol structural domains were found at high levels in late endosomes and in the trans-Golgi network, but were not found at statistically significant levels in early endosomes, lysosomes or the endoplasmic reticulum. We discuss the relevance of these results to understanding the role of lipid lateral organization in biological membranes. (C) 2011 Elsevier Inc. All rights reserved.
Original languageEnglish
Pages (from-to)21-30
Number of pages10
JournalJournal of Structural Biology
Volume175
Issue number1
DOIs
StatePublished - Jul 2011

Keywords

  • Cell membrane
  • Ceramide
  • Electron microscopy
  • Late endosomes
  • Lipid domains
  • Lipid rafts

All Science Journal Classification (ASJC) codes

  • Structural Biology

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