Assembly of proteoglycan aggregates in cultures of chondrocytes from bovine tracheal cartilage.
The assembly of proteoglycan aggregates in chondrocyte cell cultures was examined in pulse-chase experiments with the use of [35S]sulphate for labelling. Rate-zonal centrifugation in linear sucrose density gradients (10-50%, w/v) was used to separate the aggregated proteoglycans from monomers and to assess the size of the newly formed aggregates. The proportion of aggregates stabilized by link protein was assessed by competition with added exogenous aggregate components. The capacity of the proteoglycans synthesized in culture to compete with exogenous nasal-cartilage proteoglycans for binding was studied in dissociation-reassociation experiments. The results were as follows. (a) The proteoglycan monomers and the hyaluronic acid are exported separately and combined extracellularly. (b) The size of the aggregates increases gradually with time as the proportion of monomers bound to hyaluronic acid increases. (c) All of the aggregates present at a particular time appear to be link-stabilized and therefore not dissociated by added excess of nasal-cartilage proteoglycan monomer or hyaluronic acid oligomers. (d) The free monomer is apparently present as a complex with link protein. The monomer-link complexes are then aggregated to the hyaluronic acid. (e) The aggregates synthesized in vitro and the nasal-cartilage aggregates differ when tested for link-stabilization by incubation at low pH. The aggregates synthesized in vitro were completely dissociated whereas the cartilage proteoglycans remained aggregated. The results obtained from dissociation-reassociation experiments performed at low pH indicate that the proteoglycan monomer synthesized in vitro does not bind the hyaluronic acid or the link protein as strongly as does the nasal-cartilage monomer.
D000818:Animals; D002356:Cartilage / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D002501:Centrifugation, Zonal; D006820:Hyaluronic Acid / Q000378:metabolism; D046911:Macromolecular Substances; D009844:Oligosaccharides / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D014132:Trachea / Q000166:cytology / Q000378:metabolism
D046911:Macromolecular Substances; D009844:Oligosaccharides; D011509:Proteoglycans; D013431:Sulfates; D006820:Hyaluronic Acid