[HTML][HTML] Effect of ceramide N-acyl chain and polar headgroup structure on the properties of ordered lipid domains (lipid rafts)

P Sawatzki, T Kolter, R Bittman, E London - Biochimica et Biophysica …, 2007 - Elsevier
P Sawatzki, T Kolter, R Bittman, E London
Biochimica et Biophysica Acta (BBA)-Biomembranes, 2007Elsevier
Ceramides are sphingolipids that greatly stabilize ordered membrane domains (lipid rafts),
and displace cholesterol from them. Ceramide-rich rafts have been implicated in diverse
biological processes. Because ceramide analogues have been useful for probing the
biological function of ceramide, and may have biomedical applications, it is important to
characterize how ceramide structure affects membrane properties, including lipid raft
stability and composition. In this report, fluorescence quenching assays were used to …
Ceramides are sphingolipids that greatly stabilize ordered membrane domains (lipid rafts), and displace cholesterol from them. Ceramide-rich rafts have been implicated in diverse biological processes. Because ceramide analogues have been useful for probing the biological function of ceramide, and may have biomedical applications, it is important to characterize how ceramide structure affects membrane properties, including lipid raft stability and composition. In this report, fluorescence quenching assays were used to evaluate the effect of analogues of ceramide with different N-acyl chains or different sphingoid backbones on raft stability and sterol content. The effect of replacing 18 mol% of sphingomyelin (SM) with ceramide in vesicles composed of a 1:1 (mol:mol) mixture of SM and dioleoylphosphatidylcholine (DOPC), with or without 25 mol% sterol, was examined. In the absence of sterol, the thermal stability of the SM-rich ordered domains increased with ceramide N-acyl chain length in the order C2:0∼C6:0∼C8:0<no ceramide<C12:0<C16:0. In vesicles containing 25 mol% cholesterol (1:1:0.66 sphingolipid:DOPC:cholesterol), the dependence of raft stability on ceramide N-acyl chain length increased in the order C8:0∼C6:0<C2:0<C12:0∼no ceramide<C16:0. We also studied the stability of lipid rafts in the presence of N-lauroyl- and N-palmitoylsphingosine analogues containing altered structures in or near the polar portion of the sphingoid base. In almost all cases, the analogues stabilized rafts to about the same degree as a normal ceramide containing the same acyl chain. The only exception was N-palmitoyl-4D-ribophytosphingosine, which was very strongly raft-stabilizing. We conclude that variations in sphingoid base structure induce only insignificant changes in raft properties. N-Lauroyl and N-palmitoylsphingosine and their analogues displaced sterol from rafts to a significant degree. Both C12:0 and C16:0 analogues of ceramide may be good mimics of natural ceramide, and useful for cellular studies in which maintenance of the normal physical properties of ceramide are important.
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