Characterization and cDNA cloning of "glycoreceptor" on mouse neuroblastoma cells.

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We have prepared a streptavidin-based neoglycoprotein as a glycoprobe ; GT1b oligosaccharides were covalently coupled to streptavidin by reductive amination, yielding a glycosylated-streptavidin carrying 13 GT1b oligosaccharides, and then the glycosylated-streptavidin were polymerized with biotinylated-bovine serum albumin, yielding a polymer carrying more than 140 oligosaccharides. The polymer was used as a probe to detect activity of a "glycoreceptor" on the surface of primary-cultured neuron cells from hippocampus or neuroblastoma cells, NG1O8-15. The probe bound to NG1O8-15 cells with the apparent Kd value with 540 pM and the Bmax with 1200 sites/cell, suggesting the presence of high affinity receptor for the probe. We also found that addition of GT1b oligosaccharides or the probe to culture medium of the cells induced the activation of calmodulin-dependent protein kinase II (CaMKII) in the cytosol. These results suggest that neuronal cells have a glycoreceptor for GT1b oligosaccharides, stimulation of which induces CaMKII activation in the cytosol.