Demonstration of link protein in proteoglycan aggregates from human intervertebral disc.
Proteoglycan aggregates free of non-aggregating proteoglycan have been prepared from the annuli fibrosi and nuclei pulposi of intervertebral discs of three human lumbar spines by extraction with 4M-guanidinium chloride, associative density gradient centrifugation, and chromatography on Sepharose CL-2B. The aggregate (A1-2B.V0) was subjected to dissociative density-gradient ultracentrifugation. Three proteins of Mr 38 900, 44 200 and 50 100 found in the fraction of low buoyant density (A1-2B.V0-D4) reacted with antibodies to link protein from newborn human articular cartilage. After reduction with mercaptoethanol, two proteins of Mr 43 000 and two of Mr 20 000 and 14 000 were seen. The A1-2B.V0-D4 fraction, labelled with 125I, coeluted with both hyaluronate and a hyaluronate oligosaccharide (HA14) on a Sepharose CL-2B column. HA10 and HA14 reduced the viscosity of A1 fractions; HA4, HA6 and HA8 did not. HA14 decreased the viscosity of disc proteoglycans less than it did that of bovine cartilage proteoglycans. Thus, although a link protein was present in human intervertebral disc, it stabilized proteoglycan aggregates less well than did the link protein from bovine nasal cartilage.
D000368:Aged; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins; D006801:Humans; D006820:Hyaluronic Acid / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D007403:Intervertebral Disc / Q000032:analysis*; D046911:Macromolecular Substances; D008875:Middle Aged; D009844:Oligosaccharides / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D014783:Viscosity; D000099249:Proteoglycan Link Protein
D016326:Extracellular Matrix Proteins; D006820:Hyaluronic Acid; D046911:Macromolecular Substances; D009844:Oligosaccharides; D011506:Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein