In two-component phosphatidylcholine bilayers with coexisting liquidand Pβ, gel phases, the distribution between phases of low concentrations of glycosphingolipids can be determined by freeze-etch electron microscopy after labeling the glycolipid with a suitable protein. We have found that the distribution depends upon the glycosphingolipid species (Rock, P. et al., (1991) Biochemistry 30, 19-25). Using this technique with cholera toxin as the protein label and bilayers formed from dipalmitoyl- and dielaidoylphosphatidylcholine (1:1) containing <1 mol% GM1, we have studied the distribution of a family of GM1 homologues differing in the acyl chain and sphingoid base moieties. The GM1 preference for the Pβ ripple phase decreases with decreasing acyl chain length and increasing unsaturation. GM1 with either a C18:1 or C20:1 sphingoid base shows similar distributions in liquid and gel phases. When the molecules are preferentially found in the Pβ, phase, they are positioned along unique loci in both Λ and Λ/2 forms of the ripple structure. This localization and acyl chain dependence reflect the volume, shape and localization of molecular packing defects in the Pβ, phase. The ganglioside inclusions stabilize the Pβ, phase and form compositional domains of unique topography.
- Electron microscopy
- fluorescence recovery after photobleaching
- Galβ1 → 3GalNAcβ1 → 4Gal(3← 2αNeu5Ac)β1 → 4G1cβ1 → 1Cer
- high-performance liquid chromatography
- thin-layer chromatography
ASJC Scopus subject areas
- Cell Biology