Carbohydrate-binding specificity of pokeweed mitogens.
The carbohydrate-binding specificity of two pokeweed (Phytolacca americana) mitogens (Pa-1 and Pa-2) was investigated by means of hemagglutination inhibition assays and the quantitative inhibition of the binding of 125I-labeled lectins to human erythrocytes using various oligosaccharides, glycopeptides and glycoproteins as hapten inhibitors. Among the inhibitors employed in this study, chitin oligosaccharides and the glycopeptides and glycoproteins which bear sugar chains of the type found in serum glycoproteins, particularly PAS-1 glycoprotein and band-3 glycoprotein of human erythrocyte membranes, exerted strong inhibitory activity. The inhibitory constants of band-3 glycoprotein toward the binding of both mitogens to human erythrocytes were found to be very close to the association constants of the mitogens to the cells. Furthermore, the results of competitive binding studies between Pa-1 and Pa-2 indicated that these mitogens share a common oligosaccharide chains on the erythrocyte surface. To isolate the membrane receptors for these two mitogens, the solubilized membranes of human erythrocytes were subjected to affinity chromatographies using Pa-1-Sepharose 4B and Pa-2-Sepharose 4B as specific adsorbents. In both cases of these two specific adsorbents, band-3 glycoprotein was found to bind most strongly. These results suggest that two pokeweed mitogens have essentially the same carbohydrate-binding specificity and they bind primarily to the sugar chains of band-3 glycoprotein, possibly to the core structure of the sugar chains containing a di-N-acetylchitobiose moiety, on human erythrocytes.
D001667:Binding, Competitive; D050260:Carbohydrate Metabolism; D004912:Erythrocytes / Q000378:metabolism; D006020:Glycopeptides / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006241:Haptens; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000378:metabolism*; D011955:Receptors, Drug / Q000302:isolation & purification / Q000378:metabolism
D006020:Glycopeptides; D006023:Glycoproteins; D006241:Haptens; D037102:Lectins; D011955:Receptors, Drug