Glycoprotein biosynthesis by organ cultures of hamster trachea.
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.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D006224:Cricetinae; D005643:Fucose / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification; D009924:Organ Culture Techniques; D013997:Time Factors; D014132:Trachea / Q000378:metabolism*; D014574:Uronic Acids / Q000032:analysis
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D014574:Uronic Acids; D005643:Fucose; D005944:Glucosamine