Wistaria sinensis agglutinin: purification, carbohydrate specificity, and characterisation of the combining site.
A 2-acetamido-2-deoxy-D-galactose-binding agglutinin from Wistaria sinensis seeds, purified by affinity chromatography on a 2-acetamido-2-deoxy-D-galactose-starch conjugate, was homogeneous as judged by poly(acrylamide) disc gel electrophoresis. It had a mol. wt. of 66,000 (gel filtration on Sephadex G-150); on electrophoresis on SDS-poly(acrylamide) gel in the presence of 2-mercaptoethanol, it dissociated into sub-units of mol. wt. 34,000, suggesting the agglutinin to be a dimer; and it was a glycoprotein containing 4.8% of carbohydrate. It agglutinated several vertebrate erythrocytes, including human regardless of the blood group. In hapten-inhibition assays, 2-acetamido-2-deoxy-D-galactose and its glycosides were found to be better inhibitors than D-galactose and its glycosides, but N-acetyl-lactosamine was the most potent inhibitor. The binding involved HO-3,4 of the haptens and HO-2 partially.
Ahmed
H
H
Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Calcutta.
Chatterjee
B P
BP
D000818:Animals; D002241:Carbohydrates; D002846:Chromatography, Affinity; D006384:Hemagglutination; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D037102:Lectins / Q000302:isolation & purification*; D008970:Molecular Weight; D037121:Plant Lectins; D034781:Receptors, N-Acetylglucosamine; D012639:Seeds / Q000032:analysis
D002241:Carbohydrates; D037102:Lectins; D037121:Plant Lectins; D034781:Receptors, N-Acetylglucosamine; C040129:wisteria lectin