Sphingomyelin modulates the transbilayer distribution of galactosylceramide in phospholipid membranes

Peter Mattjus, Barbara Malewicz, Jacob T. Valiyaveettil, Wolfgang J. Baumann, Robert Bittman, Rhoderick E Brown

Research output: Contribution to journalArticle

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Abstract

The interrelationships among sphingolipid structure, membrane curvature, and glycosphingolipid transmembrane distribution remain poorly defined despite the emerging importance of sphingolipids in curved regions and vesicle buds of biomembranes. Here, we describe a novel approach to investigate the transmembrane distribution of galactosylceramide in phospholipid small unilamellar vesicles by 13C NMR spectroscopy. Quantitation of the transbilayer distribution of [6-13C]galactosylceramide (99.8% isotopic enrichment) was achieved by exposure of vesicles to the paramagnetic ion, Mn2+. The data show that [6-13C]galactosylceramide prefers (70%) the inner leaflet of phosphatidylcholine vesicles. Increasing the sphingomyelin content of the 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles shifted galactosylceramide from the inner to the outer leaflet. The amount of galactosylceramide localized in the inner leaflet decreased from 70% in pure 1-palmitoyl-2-oleoyl-phosphatidylcholine vesicles to only 40% in 1-palmitoyl-2-oleoyl-phosphatidylcholine/sphingomyelin (1:2) vesicles. The present study demonstrates that sphingomyelin can dramatically alter the transbilayer distribution of a monohexosylceramide, such as galactosylceramide, in 1-palmitoyl-2-oleoyl-phosphatidylcholine/sphingomyelin vesicles. The results suggest that sphingolipid-sphingolipid interactions that occur even in the absence of cholesterol play a role in controlling the transmembrane distributions of cerebrosides.

Original languageEnglish (US)
Pages (from-to)19476-19481
Number of pages6
JournalJournal of Biological Chemistry
Volume277
Issue number22
DOIs
StatePublished - May 31 2002

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