Glycosphingolipid phase behaviour in unsaturated phosphatidylcholine bilayers: a 2H-NMR study

2H-NMR was employed to consider the arrangement of a glycosphingolipid, N-(lignoceroyl-d47)galactosylceramide, in bilayers of the mono-unsaturated phospholipid, 1-stearoyl-2-oleoylphosphatidylcholine. The deuterated glycolipid prepared by partial synthesis was incorporated at concentrations ranging from 5 mol% to 53 mol% into unsonicated liposomes, and its spectra were recorded from +76 degrees C to -10 degrees C. First spectral moments were plotted as a function of temperature for each sample composition and, along with inspection of the spectra, were employed to infer a phase diagram describing glycolipid behaviour in the unsaturated phospholipid host matrix. It was possible to refine the result using 2H-NMR difference spectroscopy. The phase diagram obtained was indicative of peritectic behaviour. At glycolipid concentrations exceeding about 20 mol% there was considerable tendency to glycolipid phase separation--as indicated by coexistence of fluid phospholipid-enriched and gel phase glycolipid-enriched domains over a wide range of temperatures, and by coexistence of distinct ordered phase domains at lower temperature. In contrast, at lower glycolipid concentrations reflective of many biological membranes, the lipid components were miscible in both the liquid crystal and gel phases, with only a narrow temperature range of fluid and ordered phase coexistence. For the fluid phase at low glycolipid concentrations, spectra of the deuterated glycolipid 24-carbon fatty acid suggest that orientational order is low for a number of methylene groups near the methyl end of the chain.