Lectin from embryos and oocytes of Xenopus laevis. Purification and properties.
Soluble extracts of Xenopus laevis blastula stage embryos, oocytes, and adult liver contain lectin activities detected by agglutination of trypsinized, glutaraldehyde-fixed rabbit erythrocytes. Lectin from the embryos and oocytes was purified by affinity chromatography on a column derivatized with melibiose. Trace contaminants were removed either by preparative isoelectric focusing or by gel filtration. Based on its behavior on Sepharose 6B the purified oocyte lectin has an apparent molecular weight of approximately 480,000. On sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions there were two major bands with molecular weight ranges of about 43,000 and 45,000, with diffuse trails. Since the purified lectin contains about 20% saccharides by weight and since both bands are glycosylated, diffuseness might be due to variable glycosylation. Heterogeneity was indicated by isoelectric focusing in polyacrylamide gels, which showed four protein bands with isoelectric points ranging from 4.4 to 4.9. Lectins from both embryos and oocytes comprised about 1 to 2% of the total soluble protein and could not be distinguished by sodium dodecyl sulfate polyacrylamide gel electrophoresis. However, the specific hemagglutination activity of the purified oocyte lectin was, on the average, 7-fold higher. Levels in crude extracts of liver were 3 orders of magnitude lower than those from oocytes. The hemagglutination activities of the lectins from embryos, oocytes, and adult liver required Ca2+ and were blocked by similar concentrations of both alpha- and beta-galactosides.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001755:Blastocyst / Q000032:analysis*; D002241:Carbohydrates; D002846:Chromatography, Affinity; D004625:Embryo, Nonmammalian / Q000032:analysis*; D005260:Female; D006384:Hemagglutination; D006595:Hexosamines / Q000032:analysis; D037102:Lectins / Q000302:isolation & purification*; D008099:Liver / Q000032:analysis; D009865:Oocytes / Q000032:analysis*; D010063:Ovum / Q000032:analysis*; D014981:Xenopus
D000596:Amino Acids; D002241:Carbohydrates; D006595:Hexosamines; D037102:Lectins