Purification and characterization of novel lectins from Great Northern bean, Phaseolus vulgaris L.
Two lectins, GNL-1 and 2, were isolated from extracts of Great Northern bean powder through fractionation with ammonium sulfate, ion-exchange chromatographies on CM- and DEAE-celluloses, and gel filtration chromatography on Sephacryl S-200 HR. These lectins were shown to be homogenous by gel electrophoresis, gel filtration, and isoelectric focusing. The lectins (GNL-1 and 2) have molecular masses of 175 and 145 kDa on gel filtration, respectively. They yield three bands having the respective same molecular masses on SDS-PAGE (GNL-1; alpha-subunit of 34.5 kDa, beta of 37.0, and gamma of 39.0: GNL-2; alpha' of 34.5 kDa, beta' of 37.0, and gamma' of 39.0). Two lectins are shown to be glycoproteins and the carbohydrate contents of GNL-1 and 2 are 5.1 and 4.5%, respectively. The isoelectric points are 5.5 and 5.1 and the extinction coefficients (A 1cm 1%) at 280 nm are 11.37 and 11.45, respectively. These lectins are nonspecific in agglutination for rabbit and any types of human erythrocytes. Inhibition study shows no specificity against mono and disaccharides. On the other hand, binding assay of horseradish peroxidase-glycoproteins to the bands electroblotted onto PVDF membrane reveals that all of the subunits can bind to sugar moieties in fetuin, asialofetuin, and porcine thyroglobulin specifically. Moreover, assay of mitogenic activity shows that GNL-1 is a strong mitogen, but GNL-2 is lack of the activity.
Department of Agricultural Chemistry, Faculty of Bioresources, Mie University, Japan.
K
K
Kamemura
Y
Y
Furuichi
H
H
Umekawa
T
T
Takahashi
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D004912:Erythrocytes / Q000187:drug effects; D007887:Fabaceae / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010946:Plants, Medicinal
D000596:Amino Acids; D037102:Lectins; D037121:Plant Lectins