title: Synthesis of sulfated oligosaccharides by cystic fibrosis trachea epithelial cells.
abstract: The mucin glycoproteins in tracheal mucus of patients with cystic fibrosis is more highly sulfated than the corresponding secretions from healthy individuals [16]. In order to further characterize these differences in sulfation and possibly also glycosylation patterns, we compared the structures of sulfated mucin oligosaccharides synthesized by continuously cultured human tracheal cells transformed by simian virus 40. The synthesis of highly sulfated oligosaccharide chains in mucins secreted by normal human epithelial and submucosal cell lines were compared with mucins formed by cystic fibrosis tracheal epithelial and submucosal cell lines. The epithelial cell lines from cystic fibrosis trachea showed a higher rate of sulfate uptake and a significantly higher rate of synthesis and sulfation of high molecular weight chains. Mucins synthesized by each cell line in the presence of 35SO4 were isolated and oligosaccharide chains were released by beta-elimination and separated by ion exchange chromatography and gel filtration. The sulfated high molecular weight chains synthesized by the cystic fibrosis cell lines were characterized by methylation analysis and sequential glycosidase digestion before and after desulfation. Carbohydrate analysis yielded Fuc, Gal and GlcNAc in a ratio of 1:2:2.2 and only one galactosaminitol residue for about every 150-200 sugar residues present. The average molecular size of oligosaccharide chains in these fractions was between 30,000-40,000 daltons. These studies show that increased sulfation of oligosaccharides in mucins synthesized by cells from cystic fibrosis trachea is accompanied by a significant increase in the extension of a basic branched structure present in many of the lower molecular weight oligosaccharides.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens 30602, USA.
authors.forename: J
authors.initials: J
authors.lastname: Mendicino
authors.forename: S
authors.initials: S
authors.lastname: Sangadala
mesh_terms: D002461:Cell Line, Transformed; D002847:Chromatography, Agarose / Q000379:methods; D003550:Cystic Fibrosis / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D009077:Mucins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism; D012016:Reference Values; D013431:Sulfates / Q000737:chemistry / Q000493:pharmacokinetics; D013462:Sulfur Radioisotopes; D014132:Trachea / Q000378:metabolism*
chemical_list: D006023:Glycoproteins; D009077:Mucins; D009844:Oligosaccharides; D013431:Sulfates; D013462:Sulfur Radioisotopes; D010724:Phosphoadenosine Phosphosulfate