Isolation and characterization of a Forssman antigen-binding lectin from velvet bean (Mucuna derringiana) seeds |
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Authors: | Hanqing Mo Irwin J Goldstein |
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Institution: | (1) Department of Biological Chemistry, University of Michigan, 48109 Ann Arbor, Michigan, USA |
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Abstract: | A Forssman antigen (GalNAc 1-3GalNAc 1-3Gal 1-4Gal 1-4Glc 1-1Cer)-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 (GalNAc 1-3GalNAc)-related structures.
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, GalNAc 1-3Gal; A trisaccharide, GalNAc 1-3Fuc 1-2]Gal; Forssman disaccharide, GalNAc 1-3GalNAc; CDH (ceramide dihexoside or lactosyl ceramide) Gal 1-4Glc 1-1Cer (LacCer); CTH (ceramide trihexoside or globotriosyl ceramide), Gal 1-4Gal 1-4Glc 1-1Cer (GbOse3Cer or Gb3); globoside (globotetraosyl ceramide), GalNAc 1-3Gal 1-4Gal 1-4Glc 1-1Cer (GbOse4Cer or Gb4); Forssman antigen (globopentaosyl ceramide), GalNAc 1-3GalNAc 1-3Gal 1-4Gal 1-4Glc 1-1Cer (GbOse5Cer). |
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Keywords: | Mucuna (Mucuna derringiana) lectin velvet bean lectin haemagglutinin Forssman antigen binding lectin |
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