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  • Title: Glycoprotein biosynthesis by organ cultures of hamster trachea.
    Author: Daniel PF, Wolf G.
    Journal: Biochim Biophys Acta; 1976 Nov 18; 451(1):184-200. PubMed ID: 1009107.
    Abstract:
    Conditions have been developed for maintaining hamster tracheas in organ culture for at least 10 days. Secreted glycoproteins labelled with [14C]glucosamine and [3H]fucose were isolated from the spent medium and digested with papain, and the digest was fractionated on DEAE-Sephadex by stepwise elution with NaCl. The fractions eluted by 0.1 and 0.2 M NaCl and some of the products eluted with 0.4 M NaCl were shown to be derived from epithelial glycoproteins. Glycosaminoglycans were eluted by 0.4 M and by 1.25 M NaCl. Glycopeptides isolated from the epithelium by homogenization, ethanol precipitation and papain digestion, and defined as "intracellular", gave a very similar profile on DEAE-Sephadex. The 0.1 M glycopeptide peak was the major fraction of epithelial origin from both secreted and "intracellular" material; it labelled extensively with both glucosamine and fucose and had a molecular fraction was purified further by chromatography on Sephadex G-75 and DEAE-Sephadex; its amino acid and carbohydrate compositions were determined.
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