Abstract
Synthetic glycolipids containing an alpha-mannoside group linked by a hydrophilic spacer arm to cholesterol were incorporated into bovine erythrocytes by exchange from glycolipid-containing liposomes. When the distance between the sugar and the cholesterol moieties was approximately 26 A, functional incorporation of these glycolipids could be easily detected, as revealed by the concanavalin A-mediated agglutination of these cells. Bovine erythrocytes are not themselves susceptible to concanavalin A-mediated agglutination. The minimal concentration of concanavalin A required for agglutination of modified erythrocytes, containing 9.15 x 10(6) glycolipid molecules per cell, was 4 microgram/ml. Under these conditions, only approximately 4% of the membrane-bound cholesterol had been exchanged for the synthetic glycolipid. The observed aggregation was reversible in the presence of alpha-methyl mannoside and did not occur when beta-galactosyl-containing glycolipids were used in place of their alpha-mannoside isomers. These studies demonstrate a technique of sugar incorporation into cell membranes which should be of great advantage in studies on the roles of cell surface sugars in biological recognition. Furthermore, they demonstrate that the sugars need only be a short distance (26 A) from the membrane in order to functionally bind concanavalin A.
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