Spontaneous curvature of ganglioside GM1--effect of cross-linking.
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 Ca(2+)/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.
Department of Cell Biology, A. Mickiewicz University, Umultowska 89, PL-61614 Poznań, Poland. lumro@amu.edu.pl
Lucyna
L
Mrówczyńska
Christer
C
Lindqvist
Aleš
A
Iglič
Henry
H
Hägerstrand
D045744:Cell Line, Tumor; D002772:Cholera Toxin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000648:ultrastructure*; D005677:G(M1) Ganglioside / Q000737:chemistry*; D006801:Humans; D008856:Microscopy, Fluorescence; D009993:Osmium Tetroxide / Q000737:chemistry
D003432:Cross-Linking Reagents; D005677:G(M1) Ganglioside; D002772:Cholera Toxin; D009993:Osmium Tetroxide