A homotetrameric agglutinin with antiproliferative and mitogenic activities from haricot beans

A homotetrameric agglutinin with a molecular mass of 130 kDa was isolated from seeds of the haricot bean. The agglutinin was isolated using a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and gel filtration by fast protein liquid chromatography on Superdex 200. Haricot bean agglutinin was adsorbed on DEAE-cellulose and Affi-gel blue gel. The hemagglutinating activity of the agglutinin was stable up to 40 °C. It underwent a 40% decline when the temperature was raised to 50 °C and a complete loss when the temperature was further increased to 80 °C. The hemagglutinating activity exhibited a time-dependent loss in activity when the agglutinin was incubated at 100 °C for different durations. No activity was discernible when the agglutinin was left at 100 °C for 1 min. The activity also underwent a decline in the presence of 500 mM FeCl3 and CaCl2. Haricot bean agglutinin manifested a weaker mitogenic activity than concanavalin A toward mouse splenocytes. It exhibited antiproliferative activity toward the tumor cell lines M1 [leukemia], HepG2 [hepatoma] and L1210 [leukemia] cells.