Abstract
The lectins from seeds of Erythrina cristagalli and Erythrina corallodendron were characterized for binding to glycolipids, using a chromatogram 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 was 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 proton NMR spectroscopy it was shown to be a branched hexaglycosylceramide 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.
Highlights
Susann TenebergS, Jonas hgstrom, P e r - h eJovall, andKarl-Anders Karlsson From the Department of Medical Biochemistry, University of Gteborg, Medicinaregatan 9, S-413 90 Goteborg,Sweden
The lectins from seeds of Erythrina cristagalli and Erythrina corallodendron were characterized for binding toglycolipids, using a chromatogrambinding assay, a microtiter well assay, and glycolipids coated on erythrocytes
Plant lectins have a widespread use in glycoconjugate research and incell biology
Summary
Tination assays were performeda s described above, using a dilution of Isolation of Bovine Buttermilk Glycosphingolipids-The ganthe E . cristugulli lectin a t 0.1mg/ml. The series of intense peaks a t mlz 1591-1703 of Fig. 2 cor- In the spectrumof the permethylatedderivative (not shown), responds to the complete saccharide chain and the fatty acid, showing 4 hexoses, 2 N-acetylhexosamineas,nd a series of 16:O to 24:O non-hydroxy fatty acids This was supportedby the ions in the molecular region a t rnlz 1843-1955, which in addition indicated sphingosine as the main long-chain base. Electron Impact Mass Spectrometry of the Acid Heptaglycosylceramide-The series of peaks a t rnlz 1910-2022 of the permethylated and reducedderivative, reproduced, indicated the presence of 1 NeuAc, 4 hexoses, and 2 N-acetylhexosamines combined with a series of non-hydroxy fatty acids with 16(mlz 1910) to (mlz2022) carbon atoms.A peaka t rnlz 376 for the permethylated derivative (not shown) and a t rnlz 334 for the permethylated-reduced sample (Fig. 3) indicated a terminal NeuAc. Further sequence ions for the permethylated derivative(not reproduced)were found a t rnlz 825 (NeuAchexose-N-acetylhexosamine),a t rnlz 219 and 187(terminal hexose), rnlz 464 (terminal hexose-N-acetylhexosamine),rnlz 1478.
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