HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: M500060-JLR200v1 46/8/1678    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Yu, C. Articles by Dobrowsky, R. T. Search for Related Content PubMed PubMed Citation Articles by Yu, C. Articles by Dobrowsky, R. T. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 46, 1678-1691, August 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Ceramide displaces cholesterol from lipid rafts and decreases the association of the cholesterol binding protein caveolin-1

Cuijuan Yu, Michail Alterman and Rick T. Dobrowsky¹

Department of Pharmacology and Toxicology, University of Kansas, Lawrence, KS 66045

Published, JLR Papers in Press, May 1, 2005. DOI 10.1194/jlr.M500060-JLR200

¹ To whom correspondence should be addressed. e-mail: dobrowsky{at}ku.edu

Addition of exogenous ceramide causes a significant displacement of cholesterol in lipid raft model membranes. However, whether ceramide-induced cholesterol displacement is sufficient to alter the protein composition of caveolin-enriched lipid raft membranes is unknown. Therefore, we examined whether increasing endogenous ceramide levels with bacterial sphingomyelinase (bSMase) depleted cholesterol and changed the protein composition of caveolin-enriched membranes (CEMs) isolated from immortalized Schwann cells. bSMase increased ceramide levels severalfold and decreased the cholesterol content of detergent-insoluble CEMs by 25–50% within 2 h. To examine the effect of ceramide on the protein composition of the CEMs, we performed a quantitative proteomic analysis using stable isotope labeling of cells in culture and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Although ceramide rapidly depleted lipid raft cholesterol, the levels of the cholesterol binding protein caveolin-1 (Cav-1) decreased by 25% only after 8 h. Importantly, replenishing the cells with cholesterol rapidly reversed the loss of Cav-1 from the CEMs. Ceramide-induced cholesterol depletion increased the association of 5'-nucleotidase and ATP synthase ß-subunit with the CEMs but had a minimal effect on changing the abundance of other lipid raft proteins, such as flotillin-1 and G-proteins.

These results suggest that the ceramide-induced loss of cholesterol from CEMs may contribute to altering the lipid raft proteome.

Abbreviations: bSMase, bacterial sphingomyelinase; Cav-1, caveolin-1; CEM, caveolin-enriched membrane; dFCS, dialyzed fetal calf serum; MALDI-TOF MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry; MßCD, methyl-ß-cyclodextrin; SC, Schwann cell; SILAC, stable isotope labeling with amino acids in cell culture

Supplementary key words caveolae • proteomics • sphingolipid • Schwann cells • stable isotope labeling with amino acids in cell culture • SILAC

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. Heakal and M. Kester Nanoliposomal Short-Chain Ceramide Inhibits Agonist-Dependent Translocation of Neurotensin Receptor 1 to Structured Membrane Microdomains in Breast Cancer Cells Mol. Cancer Res., May 1, 2009; 7(5): 724 - 734. [Abstract] [Full Text] [PDF]

				R. Chan, P. D. Uchil, J. Jin, G. Shui, D. E. Ott, W. Mothes, and M. R. Wenk Retroviruses Human Immunodeficiency Virus and Murine Leukemia Virus Are Enriched in Phosphoinositides J. Virol., November 15, 2008; 82(22): 11228 - 11238. [Abstract] [Full Text] [PDF]

				F. Galbiati, V. Basso, L. Cantuti, M. I. Givogri, A. Lopez-Rosas, N. Perez, C. Vasu, H. Cao, R. van Breemen, A. Mondino, et al. Autonomic Denervation of Lymphoid Organs Leads to Epigenetic Immune Atrophy in a Mouse Model of Krabbe Disease J. Neurosci., December 12, 2007; 27(50): 13730 - 13738. [Abstract] [Full Text] [PDF]

				A. Rebillard, X. Tekpli, O. Meurette, O. Sergent, G. LeMoigne-Muller, L. Vernhet, M. Gorria, M. Chevanne, M. Christmann, B. Kaina, et al. Cisplatin-Induced Apoptosis Involves Membrane Fluidification via Inhibition of NHE1 in Human Colon Cancer Cells Cancer Res., August 15, 2007; 67(16): 7865 - 7874. [Abstract] [Full Text] [PDF]

				N. E. Furland, G. M. Oresti, S. S. Antollini, A. Venturino, E. N. Maldonado, and M. I. Aveldano Very Long-chain Polyunsaturated Fatty Acids Are the Major Acyl Groups of Sphingomyelins and Ceramides in the Head of Mammalian Spermatozoa J. Biol. Chem., June 22, 2007; 282(25): 18151 - 18161. [Abstract] [Full Text] [PDF]

				K. Krishnamurthy, S. Dasgupta, and E. Bieberich Development and characterization of a novel anti-ceramide antibody J. Lipid Res., April 1, 2007; 48(4): 968 - 975. [Abstract] [Full Text] [PDF]

				W. Chen, D. B. Jump, W. J. Esselman, and J. V. Busik Inhibition of Cytokine Signaling in Human Retinal Endothelial Cells through Modification of Caveolae/Lipid Rafts by Docosahexaenoic Acid Invest. Ophthalmol. Vis. Sci., January 1, 2007; 48(1): 18 - 26. [Abstract] [Full Text] [PDF]

				Megha, O. Bakht, and E. London Cholesterol Precursors Stabilize Ordinary and Ceramide-rich Ordered Lipid Domains (Lipid Rafts) to Different Degrees: IMPLICATIONS FOR THE BLOCH HYPOTHESIS AND STEROL BIOSYNTHESIS DISORDERS J. Biol. Chem., August 4, 2006; 281(31): 21903 - 21913. [Abstract] [Full Text] [PDF]

				S. Jean-Louis, S. Akare, M. A. Ali, E. A. Mash Jr., E. Meuillet, and J. D. Martinez Deoxycholic Acid Induces Intracellular Signaling through Membrane Perturbations J. Biol. Chem., May 26, 2006; 281(21): 14948 - 14960. [Abstract] [Full Text] [PDF]