ISOLATION AND CHARACTERIZATION OF A PHYTOHEMAGGLUTININ FROM LENTIL

A hemagglutinin from the common lentil has been isolated from crude extracts by precipitation in slightly acidified water followed by either DEAE-cellulose chromatography or elution from a Sephadex G-100 column with 0.1 m D-glucose. The hemagglutinin thus prepared exhibited a single precipitin arc on immunoelectrophoresis and a single symmetrical peak on ultracentrifugation. The sedimentation coefficient did not vary appreciably over a range of concentration from 2 to 8 mg per ml, and the S20,w0 = 4.0 S. Despite the homogeneity indicated by the above criteria, Lens culinaris hemagglutinin consistently migrated as a smear upon prolonged electrophoresis on cellulose acetate at pH 8.6. Red cells of different animal species were agglutinated to specificextents by the hemagglutinin; some strongly, others moderately, still others not at all. A saccharide binding site was implicated in the agglutination reaction by the following observations: (a) several soluble saccharides inhibited agglutination to various degrees; (b) D-glucose, a good inhibitor of agglutination, could protect the hemagglutinin against heat denaturation; (c) the hemagglutinin adhered to columns of dextran gels in which the pore size was such that the aggluti nin could reach the interior of the gel; (d) the hemagglutinin could be eluted from such gels with D-glucose but not with buffer alone. © 1969, American Chemical Society. All rights reserved.