CHARACTERIZATION OF BINDING OF GAL-BETA-4GLCNAC-SPECIFIC LECTINS FROMERYTHRINA-CRISTAGALLI AND ERYTHRINA-CORALLODENDRON TO GLYCOSPHINGOLIPIDS - DETECTION, ISOLATION, AND CHARACTERIZATION OF A NOVEL GLYCOSPHINGOLIPID OF BOVINE BUTTERMILK

The lectins from seeds of Erythrina cristagalli and Erythrina corallod endron were characterized for binding to glycolipids, using a chromato gram binding assay, a microtiter well assay, and glycolipids coated on erythrocytes. Both lectins bound to glycolipids having a terminal Gal beta 4GlcNAc sequence and also, with similar affinity, to glycolipids with terminal Fuc alpha 2Gal beta 4GlcNAc (blood group H determinant on a type 2 chain). All other substitutions of Gal beta 4GlcNAc tested abolished the binding. A binding epitope for the Erythrina lectins wa s considered by comparison of minimum energy conformations of binding and nonbinding glycolipids. A non-acid glycolipid, with lectin binding activity, was found in bovine buttermilk. By mass spectrometry and pr oton NMR spectroscopy it was shown to be a branched hexaglycosylcerami de with the structure Gal beta 4Glc-NAc beta 6(Gal beta 4GlcNAc beta 3 )Gal beta 4Glc beta Cer. This glycosphingolipid has not been reported before.