Chemical mapping of ceramide distribution in sphingomyelin-rich domains in monolayers.
The incorporation of ceramide in phase-separated monolayers of ternary lipid mixtures has been studied by a combination of atomic force microscopy (AFM), fluorescence, and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Replacement of a fraction of the sphingomyelin by ceramide in DOPC/SM/cholesterol monolayers leads to changes in the SM-cholesterol-rich liquid-ordered domains. AFM shows the formation of heterogeneous domains with small raised islands that are assigned to a ceramide-rich gel phase. ToF-SIMS provides conclusive evidence for the localization of SM and ceramide in ordered domains and shows that ceramide is heterogeneously distributed in small islands throughout the domains. The results indicate the utility of combining AFM and ToF-SIMS for understanding compositions of phase-separated membranes.
Popov
Jesse
J
Steacie Institute for Molecular Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Ontario, Canada.
Vobornik
Dusan
D
Coban
Oana
O
Keating
Eleonora
E
Miller
David
D
Francis
James
J
Petersen
Nils O
NO
Johnston
Linda J
LJ
D002518:Ceramides / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D010316:Particle Size; D013050:Spectrometry, Fluorescence; D018629:Spectrometry, Mass, Secondary Ion; D013109:Sphingomyelins / Q000737:chemistry*; D013499:Surface Properties
D002518:Ceramides; D008567:Membranes, Artificial; D013109:Sphingomyelins