Glycolipid-lectin interactions: reactivity of lectins from Helix pomatia, Wisteria floribunda, and Dolichos biflorus with glycolipids containing N-acetylgalactosamine.
The autoradiographic detection of 125I-labeled lectins binding to glycolipids on thin-layer chromatograms can be used to rapidly analyze total glycolipid extracts of cells or tissues for specific oligosaccharide structures. The Helix pomatia lectin which binds with high affinity to terminal alpha-linked GalNAc residues did not bind to globoside (terminal beta 1-3GalNAc) but did bind the ganglioside GM2 and its asialo derivative which have terminal beta 1-4GalNAc residues. The lectin from Dolichos biflorus bound specifically to the Forssman glycolipid with relatively low affinity. The lectin from Wisteria floribunda was bound to Forssman glycolipid, globoside, and the asialo derivative of the ganglioside GM2. The interactions of these lectins with the glycolipid-derived, 3H-labeled oligosaccharides was also analyzed by affinity chromatography. The results indicated that the reactivity of multivalent carbohydrate-binding proteins with polyvalent surfaces of glycolipids is strong enough to permit detection of low-affinity interactions that may not be observed in binding assays that are based on carbohydrate-protein interactions in solution. The autoradiographic analysis of 125I-Helix pomatia lectin binding to thin-layer chromatograms of total lipid extracts from human erythrocyte membranes detected the quantitative differences in the A-active glycolipids from type A1 and A2 cells.
Torres
B V
BV
Department of Biochemistry and Nutrition, Virginia Tech, Blacksburg 24061.
McCrumb
D K
DK
Smith
D F
DF
D000116:Acetylgalactosamine / Q000032:analysis*; D000818:Animals; D001345:Autoradiography; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D004912:Erythrocytes / Q000378:metabolism; D005678:G(M2) Ganglioside / Q000378:metabolism; D005688:Galactosamine / Q000031:analogs & derivatives*; D005915:Globosides / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism*; D006372:Helix, Snails; D037102:Lectins / Q000378:metabolism*; D009844:Oligosaccharides / Q000032:analysis; D012756:Sheep
D005915:Globosides; D006017:Glycolipids; D037102:Lectins; D009844:Oligosaccharides; D005678:G(M2) Ganglioside; C010163:Forssman glycolipid; D005688:Galactosamine; D000116:Acetylgalactosamine