The glycolipid specificity ofErythrina cristagalli agglutinin

Autor: Sarah Ehrlich-Rogozinski, Antonio De Maio, Nathan Sharon, Halina Lis
Rok vydání: 1987
Předmět:
Zdroj: Glycoconjugate Journal. 4:379-390
ISSN: 1573-4986
0282-0080
DOI: 10.1007/bf01048371
Popis: The interaction of125I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with125I-ECA. Afterin situ treatment of the chromatograms with α-galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Galα3Galβ4GlcNAcβ3Galβ4Glcβ1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.
Databáze: OpenAIRE
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