Isolation and Characterization of Rana catesbeiana Lectin and Demonstration of the Lectin-binding Glycoprotein of Rodent and Human Tumor Cell Membranes1
نویسندگان
چکیده
A lectin isolated from Rana catesbeiana eggs preferentially aggluti nates a large variety of human and animal tumor cells but not normal red blood cells, lymphocytes, or fibroblasts. The phenomenon correlates with a higher binding activity of the lectin with tumor cells. Chemical and physical analysis of the purified lectin indicates that the lectin is a low molecular weight basic polypeptide with five intrachain disulfide bonds. Its agglutination of tumor cells was abolished by blocking the amino group. The lectin strongly binds with a large variety of tumor cells but binds only minimally with fibroblasts, lymphocytes, and erythrocytes. Tumor cell agglutination induced by this lectin was strongly inhibited by submaxillary mucin, to a lesser degree by fetuin and keratan sulfate, and not at all by less-sialylated glycoproteins, such as transferrin. Inhibition by mucin or fetuin was greatly reduced by desialylation of glycoprotein with sialidase. Treatment of tumor cells with sialidase greatly reduced the lectin-dependent agglutination, and the sialidase-dependent reduction of tumor cell agglutination was inhibited by the sialidase inhibitor 2,3dehydro-2-deoxy-jY-acetylneuraminic acid. However, tumor cell agglutin ation was not inhibited by chondroitin sulfates or hyaluronic acid. Thus, the lectin-dependent tumor cell agglutination is due to a high density of sialic acid at the cell surface. The receptor glycoprotein that interacts with this lectin was demonstrated in the detergent-insoluble fraction of a variety of tumor cells by sodium dodecyl sulfate:polyacrylamide gel electrophoresis, followed by Western blotting with lectin and anti-lectin antibodies. The presence of a common high molecular weight lectinbinding glycoprotein in various tumor cells was demonstrated.
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