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Cell-free biosynthesis of erythroglycan in a microsomal fraction from K-562 cells.
Authors:T Z Russin  R A Laine  and S J Turco
Abstract:Particulate membrane preparations from K-562 human CML (chronic-myelogenous-leukaemia)-derived] cells catalyse 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 (mol.wt. 7000--10 000) called 'erythroglycan'. Incorporation of either labelled sugar increased up to 60 min of incubation time. The labelled 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 four products: (1) a disaccharide with the sequence beta GlcNAc-beta Gal; (2) a trisaccharide with the sequence betaGal-betaGlcNAc-beta Gal; (3) a larger oligosaccharide containing galactose and N-acetylglucosamine; and (4) a putative protein-linkage region.
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