Selective aggregation of proteoglycans with hyaluronic acid.
Conditions were established to separate proteoglycan aggregate (AH1) from a bovine nasal septum extract by associative rate zonal sedimentation on a NaCl gradient. The AH1 has a higher protein content than the mixed aggregate-monomer (A1) isolated by conventional associative CsCl density gradient centrifugation from a portion of the same extract. The same associative rate zonal conditions separated the A1 fraction into aggregated AH1 containing hyaluronic acid and nonaggregated proteoglycan monomer (N1) essentially free of hyaluronic acid. The AH1 fraction is richer in protein and keratin sulfate than is N1. Dissociative rate zonal sedimentation of A1 under conditions which totally sedimented most of the disaggregated monomer (AH1-D1) and the nonaggregated monomer N1 separated a less sedimentable protein and keratan sulfate-rich proteoglycan monomer (AH1-D2). Chromatography on Sepharose 2B under dissociative conditions demonstrated that the nonaggregated N1 monomer is intermediate in size between the disaggregated monomers AH1-D1 and AH1-D2. N1 has a buoyant density higher than AH1 and is practically equivalent to AH1-D1. All are dense fractions so that separation by CsCl density gradient equilibration is not feasible.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002501:Centrifugation, Zonal; D006820:Hyaluronic Acid / Q000302:isolation & purification; D007632:Keratan Sulfate; D046911:Macromolecular Substances; D009300:Nasal Septum / Q000032:analysis; D011509:Proteoglycans / Q000302:isolation & purification
D000596:Amino Acids; D046911:Macromolecular Substances; D011509:Proteoglycans; D006820:Hyaluronic Acid; D007632:Keratan Sulfate