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Isolation and characterization of a Forssman antigen-binding lectin from velvet bean (Mucuna derringiana) seeds

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Abstract

A Forssman antigen (GalNAcα1-3GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer)-binding lectin has been purified from velvet bean (Mucuna derringiana) seeds by a combination of affinity chromatography and reversed phase HPLC. This lectin agglutinates both native and trypsin-treated sheep erythrocytes as well as trypsinized rabbit erythrocytes, but neither native rabbit nor human erythrocytes, irrespective of blood group type. SDS-PAGE and gel filtration chromatography reveal the lectin to be a homodimer consisting of two 54 kDa subunits linked by non-covalent bonds. The results obtained by quantitative precipitation, haemagglutination inhibition and TLC overlay assays indicate that theMucuna lectin specifically recognizes Forssman antigen and Forssman disaccharide (GalNAcα1-3GalNAc)-related structures.

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Authors and Affiliations

  1. Department of Biological Chemistry, University of Michigan, 48109, Ann Arbor, Michigan, USA

    Hanqing Mo & Irwin J. Goldstein

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  1. Hanqing Mo
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  2. Irwin J. Goldstein
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Additional information

Abbreviations: The abbreviations and the trivial names used are: AH, 6-aminohexyl; BSA, bovine serum albumin; Cer, ceramide; HPLC, high performance liquid chromatography; PAGE, polyacrylamide gel electrophoresis; PBS, 10mm phosphate-buffered saline, pH 7,2, containing 0.15m NaCl; PMSF, phenyl methyl sulfonyl fluoride; SDS, sodium dodecyl sulphate; TFA, trifluoroacetic acid; TBS, 20mm tris-buffered saline, pH 7.2; TLC, thin-layer chromatography; A disaccharide, GalNAcα1-3Gal; A trisaccharide, GalNAcα1-3[Fucα1-2]Gal; Forssman disaccharide, GalNAcα1-3GalNAc; CDH (ceramide dihexoside or lactosyl ceramide) Galβ1-4Glcβ1-1Cer (LacCer); CTH (ceramide trihexoside or globotriosyl ceramide), Galα1-4Galβ1-4Glcβ1-1Cer (GbOse3Cer or Gb3); globoside (globotetraosyl ceramide), GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer (GbOse4Cer or Gb4); Forssman antigen (globopentaosyl ceramide), GalNAcα1-3GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer (GbOse5Cer).

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Mo, H., Goldstein, I.J. Isolation and characterization of a Forssman antigen-binding lectin from velvet bean (Mucuna derringiana) seeds. Glycoconjugate J 11, 424–431 (1994). https://doi.org/10.1007/BF00731278

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  • DOI: https://doi.org/10.1007/BF00731278

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