Properties of the lectin from the hog peanut (Amphicarpaea bracteata)

Abstract

An N-acetyl--galactosamine-specific lectin has been isolated from the two seed forms of the hog peanut (Amphicarpaea bracteata) using an affinity support containing the synthetic type A blood group trisaccharide [alpha]--GalNAc-(1,3)-[[alpha]--Fuc-(1,2)]-[beta]--Gal (Synsorb A). The affinity-purified lectin appears to be identical in both seed types. Gel filtration on Sephadex G-200 gives a single symmetrical peak corresponding to Mr 135,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows four subunit forms, each of which contains carbohydrate. Limited amino terminal sequencing indicates heterogeneity in two of the first 10 residues. The lectin contains no cysteine. There are four equivalent, noninteracting GalNAc binding sites per 135,000-Da molecule, having an association constant for methyl N-acetyl-[alpha]--galactosaminide of 4.0 x 104 -1. Precipitin and hapten inhibition studies show the lectin to be specific for terminal, nonreducing -GalNAc units, with a preference for the [alpha]-anomer and enhanced specificity for the disaccharide, GalNAc[alpha]1,3GalNAc. There is also a single adenine binding site per Mr 135,000 lectin molecule with an association constant of 1.3 x 106 -1.