Studies on extractable and resistant proteoglycans from metaphyseal and cortical bone and cartilage.
Soluble proteoglycans (SPG) were extracted from bovine (BCC) and human (HCC) costal cartilages by the dissociative method using 4 M guanidinium chloride (GuHCl). Proteoglycans which are resistant to extraction (RPG) were obtained following collagenase digestion or hydroxylamine treatment of the cartilage residues. Similarly, SPG were extracted from bovine metaphyseal and cortical bone using EDTA. The RPG were extracted from the bones using hydroxylamine. Density gradient fractionation under dissociative conditions of cartilage SPG and RPG followed by chromatography on Sepharose 2B revealed that A1D1 RPG are smaller than the SPG. SPG reacted with either collagenase or hydroxylamine are also smaller than the parent SPG. A1D1 fractions obtained from BCC-SPG and RPG or from mixtures of SPG and acid-soluble collagen are free of hydroxyproline. Hydroxyproline is not completely separated from HCC-RPG. Density gradient fractionation of bone proteoglycans and Sepharose chromatography of the A1 and A1D1 fractions showed that those obtained from metaphysis are larger than those from cortical bone. This was attributed to the presence of calcified cartilage in metaphyseal bone. The A1D1 fractions of the metaphyseal proteoglycans seemed to undergo self-association since this fraction is larger than the A1 fraction from which it is derived. Cortical bone proteoglycans do not behave similarly. Density gradient purification under dissociative conditions failed to separate hydroxyproline from the proteoglycans obtained from bone. It is hypothesized that in bone proteoglycans and collagen might be linked.
D000818:Animals; D001842:Bone and Bones / Q000032:analysis*; D002241:Carbohydrates / Q000032:analysis; D002356:Cartilage / Q000032:analysis*; D002417:Cattle; D005260:Female; D005333:Fetus; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D006909:Hydroxyproline / Q000032:analysis; D003012:Microbial Collagenase; D011247:Pregnancy; D011509:Proteoglycans / Q000302:isolation & purification*
D002241:Carbohydrates; D006146:Guanidines; D011509:Proteoglycans; D003012:Microbial Collagenase; D019791:Guanidine; D006909:Hydroxyproline