Spontaneous curvature of ganglioside GM1 – Effect of cross-linking

► Ganglioside GM1 possesses flexible spontaneous curvature (SC). ► GM1–CTB–antiCTB complex has non-flexible high overall positive SC. ► Osmium tetroxide immobilises GM1. The membrane-curvature dependent lateral distribution of outer leaflet ganglioside GM1 (GM1) and the influence of GM1 cross-linking induced by fluorophore-tagged cholera toxin subunit B (CTB) plus anti-CTB was analysed in cell membranes by fluorescence microscopy. Data are presented indicating that cross-linked GM1–ligand patches accumulated at the tips of human erythrocyte echinocytic spiculae induced by Ca2+/ionophore A23187. However, when lipid fixative osmium tetroxide was added prior to the ligand no accumulation in spiculae occurred. GM1-staining remained here distributed over the spheroid cell body and in spiculae. Similarly, osmium tetroxide completely prohibited CTB plus anti-CTB-induced GM1 patching in representatives for flat membrane, i.e. discoid erythrocytes and K562 cells. Our results demonstrate that GM1 per se shows low membrane curvature dependent distribution and therefore holds flexible spontaneous curvature. In contrast, the cross-linked GM1–ligand complex has a strong preference for highly outward curved membrane and possesses overall positive spontaneous curvature. Osmium tetroxide efficiently immobilises GM1.