Role of proteoglycans in endochondral ossification: Inhibition of calcification
Tóm tắt
Proteoglycans from bovine nasal septa and from the Swarm rat chondrosarcoma were isolated as aggregates (PGC) and as monomers (PGS). Portions of the PGC preparations were degraded with cathepsin D or chondroitinase AC. Chondroitin sulfates were isolated by differential precipitation from alkaline digests of the PGS from bovine nasal septa. The effects of these preparations at concentrations up to 2 mg/ml on the precipitation of tricalcium phosphatein vitro at pH 7.8 in 16 hours at 25°C were ascertained. To this end, the amounts of calcium and phosphate in the precipitates and in the supernates were determined. The PGC preparations were found to be very effective inhibitors; in the presence of 2 mg/ml, precipitate did not form. The PGS preparations were less effective than the PGC preparations; in the presence of 2 mg/ml, about 20% as much calcium phosphate precipitated as in their absence. The chondroitinase AC-degraded preparations at concentrations equivalent to 2 mg/ml of the PGC preparations were approximately as effective as the PGS preparations. On the other hand, the cathepsin D-degraded PGC preparations and the chondroitin sulfate chains were relatively poor inhibitors. Although the viscosity of the solutions may have influenced the rate at which the precipitates settled to the bottom of the bubes, the amounts of the tricalcium phosphate formed were related to the composition and concentration of the proteoglycan preparations.
Tài liệu tham khảo
Logan MA (1935) Composition of cartilage, bone, dentin and enamel. J Biol Chem 110:375–389
Dziewiatkowski DD (1951) Radioautographic visualization of sulfur-35 disposition in the articular cartilage and bone of suckling rats following injection of labeled sodium sulfate. J Exp Med 93:451–458
Dziewiatkowski DD (1952) Radioautographic studies of sulfate-sulfur (S35) metabolism in the articular cartilage and bone of suckling rats. J Exp Med 95:489–496
Dziewiatkowski DD, Bronner F, DiFerrante N, Archibald RM (1957) Some aspects of the metabolism of sulfate-S35 and calcium-45 in the metaphyses of immature rats. Influence of β-estradiol benzoate. J Biophys Biochem Cytol 3:151–160
Campo RD, Dziewiatkowski DD (1963) Turnover of the organic matrix of cartilage and bone as visualized by autoradiography. J Cell Biol 18:19–29
Weatherell JA, Weidmann SM (1963) The distribution of organically bound sulphate in bone and cartilage during calcification. Biochem J 89:265–267
Hirschman A, Dziewiatkowski DD (1966) Protein-polysaccharide loss during endochondral ossification: Immunochemical evidence. Science 154:393–395
Campo RD, Tourtellotte CD (1967) The composition of bovine cartilage and bone. Biochim Biophys Acta 141:614–624
Schenk RK, Spiro D, Wiener J (1967) Cartilage resorption in the tibial epiphyseal plate of growing rats. J Cell Biol 34:275–291
Matukas VJ, Krikos GA (1968) Evidence for changes in protein polysaccharide associated with the onset of calcification in cartilage. J Cell Biol 39:43–48
Scherft JP, Moskalewski S (1984) The amount of proteoglycans in cartilage matrix and the onset of mineralization. Metab Bone Dis Rel Res 5:195–203
Rubin PS, Howard JE (1950) Histochemical studies on the role of acid muco-polysaccharides in calcificbility and calcification. Trans 2nd Josiah Macy Conf Metab Interrelat 155–166
Poole AR, Pidoux I, Rosenberg L (1982) Role of proteoglycans in endochondral ossification: Immunofluorescent localization of link protein and proteoglycan monomer in bovine fetal epiphyseal growth plate. J Cell Biol 92:249–260
Howell DS, Pita JC, Marquez JF, Gatter RA (1969) Demonstration of macromolecular inhibitor(s) of calcification and nucleational factor(s) in fluid from calcifying sites in cartilage. J Clin Invest 48:630–641
Cuervo LA, Pita JC, Howell DS (1973) Inhibition of calcium phosphate mineral growth by proteoglycan aggregate fractions in a synthetic lymph. Calc Tissue Res 13:1–10
Chen CC, Boskey AL, Rosenberg LC (1984) The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth. Calc Tissue Int 36:285–290
Weinstein H, Sacks CR, Schubert M (1963) Protein-polysaccharide in connective tissue: Inhibition of phase separation. Science 142:1073–1075
Schubert M, Pras M (1968) Ground substance protein polysaccharides and the precipitation of calcium phosphate. Clin Orthop Relat Res 60:235–225
Pita JC, Cuervo LA, Madruga JE, Muller FJ, Hwell DS (1970) Evidence for a role of protein-polysaccharides in regulation of mineral phase separation in calcifying cartilage. J Clin Invest 49:2188–2197
Blumenthal NC, Posner AS, Silverman LD, Rosenberg LC (1979) Effect of proteoglycans onin vitro hydroxyapatite formation. Calc Tissue Int 27:75–82
Oegema TR, Hascall VC, Dziewiatkowski DD (1975) Isolation and characterization of proteoglycans from the Swarm rat chondrosarcoma. J Biol Chem 250:6151–6159
Barrett AJ (1970) Cathepsin D. Purification of isozymes from human and chicken liver. Biochem J 117:601–607
Dziewiatkowski DD, Tourtellotte CD, Campo RD (1968) Degradation of protein-polysaccharide (chondromucoprotein) by an enzyme extracted from cartilage. In: Quintarelli G (ed) The chemical physiology of mucopolysaccharides, Little, Brown, Boston, pp 63–79
Dziewiatkowski DD, Hascall VC, Sajdera S (1970) The effect of cathepsins from varied sources on proteinpolysaccharide (PP-L) of bovine costal cartilage and on proteoglycan subunit (PGS) of bovine nasal cartilage. Calc Tissue Res, Suppl to Vol 4:54–65
Saito H, Yamagata T, Suzuki S (1968) Enzymatic methods for the determination of small quantities of isomeric chondroitin sulfates. J Biol Chem 243:1536–1542
Hascall VC, Riolo RL, Hayward J Jr, Reynolds CC (1972) Treatment of bovine nasal cartilage proteoglycan with chondroitinases fromFlavobacterium heparinum andProteus vulgaris. J Biol Chem 247:4521–4528
Hascall VC, Riolo RL (1972) Characteristics of the proteinkeratan sulfate core and of keratan sulfate prepared from bovine nasal cartilage proteoglycan. J Biol Chem 247:4529–4538
Cuervo LA, Pita JC, Howell DS (1971) Ultramicroanalysis of pH, Pco2 and carbonic anhydrase activity at calcifying sites in cartilage. Calc Tissue Res 7:220–231
Analytical methods for atomic absorption spectrophotometry (1971) Perkin-Elmer Co., Norwalk, Conn
Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphate Anal Chem 28:1756–1759
Bitter T, Muir H (1962) A modified uronic acid carbazole reaction. Anal Biochem 4:330–334
Woodward C, Davidson EA (1968) Structure-function relationships of proteinpolysaccharide complexes: Specific ionbinding properties. Proc Nat Acad Sci 60:201–205
Martin JH and Mathews JL (1969) Mitochondral granules in chondrocytes. Calc Tissue Res 3:184–193
Anderson HC (1969) Vesicles associated with calcification in the matrix of epiphyseal cartilage. J Cell Biol 41:59–72
Anderson HC (1967) Electron microscopic studies of induced cartilage development and calcification. J Cell Biol 35:81–101
Bonucci E (1967) Fine structure of early cartilage calcification. J Ultrastruct Res 20:33–50