Cell-free biosynthesis of erythroglycan in a microsomal fraction from K-562 cells

Abstract
Particulate membrane preparations from K-562 [human chronic myelogenous leukemia-derived] cells catalyze the transfer of [3H]galactose from UDP-[3H]galactose and [3H]N-acetylglucosamine from UDP-[3H]N-acetylglucosamine into an endogenous product that on digestion with Pronase yields long-chain glycopeptides (MW 700-10,000) called erythroglycan. Incorporation of either labeled sugar increased up to 60 min of incubation time. The labeled erythroglycan was isolated by chromatography on Sephadex G-50 and characterized by digestion with endo-.beta.-galactosidase from Escherichia freundii, followed by analysis on Bio-Gel P-2 and paper chromatography. This digestion gave the following 4 products: a disaccharide with the sequence .beta.GlcNAc-.beta.Gal; a trisaccharide with the sequence .beta.Gal-.beta.GlcNAc-.beta.Gal; a larger oligosaccharide containing galactose and N-acetylglucosamine; and a putative protein-linkage region.

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