Structural characterization of human alveolar bone proteoglycans.
Proteoglycans were extracted from EDTA-demineralized human alveolar bone under dissociative conditions using 4 M guanidinium chloride in the presence of protease inhibitors. The extract was further purified by anion-exchange chromatography on DEAE-Sephacel, using a step-wise salt gradient. The proteoglycan-rich fraction was analysed for carbohydrate, protein and amino acid composition and molecular size by SDS-PAGE. Glycosaminoglycan content was determined by cellulose acetate electrophoresis after proteolysis. The sulphate isomers of the glycosaminoglycans were confirmed by Fourier-transformed infra-red spectroscopy. Two chondroitin sulphate-proteoglycan species were identified with molecular weights of 79 and 55-65 kDa, respectively. The core proteins had molecular weights of 49 kDa for both proteoglycans, with the amino acid content rich in glycine, leucine, glutamate and aspartate. The chondroitin sulphate chains were mainly as the 4-sulphate isomer forms although low but detectable amounts of 6-sulphate isomer were also present.
Department of Basic Dental Science, Dental School, University of Wales College of Medicine, Heath Park, Cardiff, U.K.
R J
RJ
Waddington
G
G
Embery
D000539:Alveolar Process / Q000737:chemistry*; D001224:Aspartic Acid / Q000032:analysis; D001857:Bone Matrix / Q000737:chemistry; D002809:Chondroitin Sulfates / Q000032:analysis; D002852:Chromatography, Ion Exchange; D004588:Electrophoresis, Cellulose Acetate; D004591:Electrophoresis, Polyacrylamide Gel; D005583:Fourier Analysis; D005971:Glutamates / Q000032:analysis; D018698:Glutamic Acid; D005998:Glycine / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis; D006595:Hexosamines / Q000032:analysis; D006601:Hexoses / Q000032:analysis; D006801:Humans; D007930:Leucine / Q000032:analysis; D008970:Molecular Weight; D011509:Proteoglycans / Q000032:analysis* / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate; D013055:Spectrophotometry, Infrared; D014574:Uronic Acids / Q000032:analysis
D005971:Glutamates; D006025:Glycosaminoglycans; D006595:Hexosamines; D006601:Hexoses; D011509:Proteoglycans; D014574:Uronic Acids; D001224:Aspartic Acid; D012967:Sodium Dodecyl Sulfate; D018698:Glutamic Acid; D002809:Chondroitin Sulfates; D007930:Leucine; D005998:Glycine