Immunochemical studies on the combining site of the blood group A-specific lima bean lectin.
The combining site of the lima bean (Phaseolus lunatus) lectin (LBL) was studied by quantitative precipitin and precipitin-inhibition assays. The lectin precipitated best with hog gastric mucosa and human ovarian cyst blood group A1 substances and moderately with A2 substances. B substances precipitated very poorly and H, Lea, Leb, and precursor I substances did not react. Blood group A1 and A2 substances reacted to varying extents and these differences are attributable to heterogeneity resulting from incomplete biosynthesis of carbohydrate chains. By inhibition of precipitation of LBL with A1 blood group substance, the lectin was found to be most specific for fucose-containing oligosaccharides having the A trisaccharide, DGalNAc alpha 1----3[L-Fuc alpha 1----2]DGal determinant. The best inhibitor, an A-specific hexasaccharide, DGalNAc alpha 1----3[LFuc alpha 1----2]DGal beta 1----3DGlcNAc beta 1----3-DGal beta 1----4DGlc, was 11 times more active than the A trisaccharide. A difucosyl oligosaccharide with a second fucose linked alpha 1----3 to the DGlcNAc is less active; fucose linked alpha 1----4 to DGlcNAc was completely inactive. These results suggest that specific interactions with the subterminal sugars may be important in the binding, and that the specificity of the lectin combining site involves at least the nonreducing terminal four and probably five sugars of the hexasaccharide. Thus LBL has a more-extended binding site than was inferred previously and is in the upper range of antibody combining-site sizes.
D000017:ABO Blood-Group System; D000818:Animals; D001665:Binding Sites; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates; D005260:Female; D005753:Gastric Mucosa / Q000276:immunology; D006801:Humans; D037102:Lectins; D009844:Oligosaccharides; D010048:Ovarian Cysts / Q000276:immunology; D037121:Plant Lectins; D013552:Swine
D000017:ABO Blood-Group System; D002241:Carbohydrates; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins; C031887:lima bean lectin