Localization of globoside and Forssman glycolipids on erythrocyte membranes.
Using the freeze-etch technique, the membrane localization of globoside, a principal glycolipid in human erythrocytes, and Forssman antigen, the chief glycolipid in sheep erythrocytes was evaluated using ferritin and colloidal gold as morphological markers for rabbit antibodies prepared against these glycolipids. Brief trypsinization of human red cell ghosts markedly aggregated intramembranous particles and permitted labeling of globoside, which appeared in a clustered arrangement. The aggregates of ferritin-anti-globoside differed from those of ferritin-wheat germ agglutinin, a label for glycophorin, which corresponded with the aggregates of intramembranous particles. Double-labeling of human trypsinized ghosts with anti-globoside/ Staphylococcal protein A-colloidal gold and ferritin-wheat germ agglutinin indicated that the patterns of labeling were different and that the aggregates of globoside did not bear a direct relationship to the intramembranous particles, which represent transmembrane proteins. Resealed sheep erythrocyte ghosts labeled with ferritin-conjugated rabbit anti-Forssman showed small clusters of Forssman glycolipid on the erythrocyte surface, which could be markedly aggregated with a second goat anti-rabbit antibody, indicating relative mobility of the small glycolipid domains. The distribution of ferritin-anti-Forssman label in sheep ghosts treated at pH 5.5 to aggregate intramembranous particles also did not show definite correspondence between intramembranous particles and the clusters of ferritin-anti-Forssman.
D015478:Antigens, Heterophile / Q000032:analysis*; D004910:Erythrocyte Membrane / Q000032:analysis / Q000276:immunology / Q000648:ultrastructure*; D004912:Erythrocytes / Q000648:ultrastructure*; D005577:Forssman Antigen / Q000032:analysis*; D005613:Freeze Etching; D005915:Globosides / Q000032:analysis* / Q000097:blood*; D006028:Glycosphingolipids / Q000032:analysis* / Q000097:blood*; D006801:Humans; D008854:Microscopy, Electron
D015478:Antigens, Heterophile; D005915:Globosides; D006028:Glycosphingolipids; C010163:Forssman glycolipid; D005577:Forssman Antigen