The chemical constitution of the proteoglycan of human intervertebral disc

Proteoglycan was prepared from three pools of normal human intervertebral discs by extraction with buffered 4M-guanidinium chloride followed by CsCl-density-gradient ultracentrifugation. Chromatography on agarose (Bio-Gel A-150m) and on DEAE-cellulose suggested a single polydisperse proteoglycan spe...

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Bibliographic Details
Published in:Biochemical journal 1976-09, Vol.157 (3), p.753-763
Main Authors: Pearce, R H, Grimmer, B J
Format: Article
Language:English
Subjects:
Adult
Chondroitin Sulfates - analysis
Glycosaminoglycans - isolation & purification
Hexosamines - analysis
Hexoses - analysis
Hexuronic Acids - analysis
Humans
Intervertebral Disc - analysis
Keratan Sulfate - analysis
Male
Middle Aged
Molecular Weight
Nitrogen - analysis
Proteoglycans - analysis
Viscosity
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Summary:Proteoglycan was prepared from three pools of normal human intervertebral discs by extraction with buffered 4M-guanidinium chloride followed by CsCl-density-gradient ultracentrifugation. Chromatography on agarose (Bio-Gel A-150m) and on DEAE-cellulose suggested a single polydisperse proteoglycan species. The intrinsic viscosities of three preparations were 166, 122 and 168 ml/g. After degradation with 0.5M-KOH containing 0.02M-NaBH4, the glycosaminoglycans were recovered quantitatively and their Ca2+ salts separated into a hexuronate-rich fraction (fraction 1), which was precipitated in 0-45% (v/v) ethanol, and a hexose-rich fraction (fraction2), which was precipitated in 45-70% (v/v) ethanol. Qualitative and quantitative analyses of the glycosaminoglycans revealed fraction 1 to be chondroitin sulphate, and fraction 2 to be keratan sulphate; the latter was contaminated with protein and possibly a small amount of another glycosaminoglycan. For both glycosaminoglycans, plots of log(mol.wt.) against weight fell close to a normal distribution. The mode for chondroitin sulphate was close to 20000; that for keratan sulphate, 10000. A threefold range of molecular weight included the central 16-84% [+/- 1 S.D. of log(mol.wt.)] of the weight of both fractions.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj1570753