title: Heterogeneity of protein-polysaccharides of porcine articular cartilage. The chondroitin sulphate proteins associaterd with collagen.
abstract: Pig articular cartilage, from which protein-polysaccharides soluble in iso-osmotic sodium acetate had been removed, was extracted in three further stages with 8m-urea in 2m-sodium acetate and with tris-HCl buffer after bacterial collagenase digestion, followed by the same urea-sodium acetate solution, thus leaving only 2% of the original uronic acid in the tissue. The histological appearance of the cartilage was unaltered until after collagenase digestion. The collagenase used did not affect the viscosity or molecular size of a protein-polysaccharide preparation obtained previously. The protein-polysaccharides in each extract differed in size, amino acid composition and protein content, but protein and keratan sulphate contents were not related to hydrodynamic size, in contrast with protein-polysaccharides extracted previously before collagenase digestion. Hydroxyproline could not be removed from those obtained by the first urea-sodium acetate extraction until degraded by heat. The galactosamine/pentose molar ratio agreed closely with the galactosamine/serine molar ratio that was destroyed on treatment with 0.5m-sodium hydroxide, showing that chondroitin sulphate was attached only to serine residues. From these molar ratios the chondroitin sulphate chains were calculated to be of the same average length in protein-polysaccharides in all three extracts although somewhat shorter than in protein-polysaccharides extracted previously. Some threonine residues were also destroyed on alkali treatment suggesting that keratan sulphate may be attached to threonine. These findings together with previous results show that differences in size, composition and physical state extend to all the protein-polysaccharides in cartilage.
mesh_terms: D000085:Acetates; D000818:Animals; D002358:Cartilage, Articular / Q000032:analysis*; D002807:Chondroitin / Q000032:analysis*; D003094:Collagen / Q000032:analysis*; D005688:Galactosamine / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006025:Glycosaminoglycans / Q000032:analysis; D006595:Hexosamines / Q000032:analysis; D006601:Hexoses / Q000032:analysis; D006909:Hydroxyproline / Q000032:analysis; D003012:Microbial Collagenase; D010429:Pentoses / Q000032:analysis; D012694:Serine / Q000032:analysis; D013431:Sulfates / Q000032:analysis; D013552:Swine; D013912:Threonine / Q000032:analysis; D014508:Urea; D014574:Uronic Acids / Q000032:analysis
chemical_list: D000085:Acetates; D006023:Glycoproteins; D006025:Glycosaminoglycans; D006595:Hexosamines; D006601:Hexoses; D010429:Pentoses; D013431:Sulfates; D014574:Uronic Acids; D013912:Threonine; D012694:Serine; D005688:Galactosamine; D014508:Urea; D002807:Chondroitin; D003094:Collagen; D003012:Microbial Collagenase; D006909:Hydroxyproline