Different sphingolipids show differential partitioning into sphingolipid/cholesterol-rich domains in lipid bilayers.
Two fluorescence-based approaches have been applied to examine the differential partitioning of fluorescent phospho- and sphingolipid molecules into sphingolipid-enriched domains modeling membrane "lipid rafts." Fluorescence-quenching measurements reveal that N-(diphenylhexatrienyl)propionyl- (DPH3:0-)-labeled gluco- and galactocerebroside partition into sphingolipid-enriched domains in sphingolipid/phosphatidylcholine/cholesterol bilayers with substantially higher affinity than do analogous sphingomyelin, ceramide, or phosphatidylcholine molecules. By contrast, the affinity of sphingomyelin and ceramide for such domains is only marginally greater than that of a phosphatidylcholine with similar hydrocarbon chains. By using direct measurements of molecular partitioning between vesicles of different compositions, we show that the relative affinities of different C(6)-NBD- and C(5)-Bodipy-labeled sphingolipids for sphingolipid-enriched domains are quantitatively, and in most circumstances even qualitatively, quite different from those found for species whose N-acyl chains more closely resemble the long saturated chains of cellular sphingolipids. These findings lend support in principle to previous suggestions that differential partitioning of different sphingolipids into "raft" domains could contribute to the differential trafficking of these species in eukaryotic cells. However, our findings also indicate that short-chain sphingolipid probes previously used to examine this phenomenon are in general ill-suited for such applications.
Wang
T Y
TY
Department of Biochemistry, McGill University, Montréal, Québec H3G 1Y6, Canada.
Silvius
J R
JR
D002784:Cholesterol / Q000737:chemistry*; D005699:Galactosylceramides / Q000737:chemistry*; D005963:Glucosylceramides / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013107:Sphingolipids / Q000737:chemistry*; D013329:Structure-Activity Relationship
D005699:Galactosylceramides; D005963:Glucosylceramides; D008051:Lipid Bilayers; D013107:Sphingolipids; D002784:Cholesterol