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. 1970 Dec;49(12):2188–2197. doi: 10.1172/JCI106437

Evidence for a role of proteinpolysaccharides in regulation of mineral phase separation in calcifying cartilage

Julio C Pita 1, Leon A Cuervo 1, Juan E Madruga 1, Francisco J Muller 1, David S Howell 1
PMCID: PMC322719  PMID: 4921198

Abstract

Our previous studies have indicated the presence of a macromolecular inhibitor of in vitro mineral growth, as well as a mineral nucleational agent in extracellular matrix fluid aspirated by micropuncture methods from epiphyseal hypertrophic cell cartilage. In this report, new miniaturized methods were used to extract proteinpolysaccharide complexes (PPC) from cartilage, to isolate a light fraction (PPL-C), and further, to separate it into R1, R2, and SR2 subfractions. These methods were applied to PPL-C complexes separated from microdissected epiphyseal cartilages and to cetylpyridinium chloride (CPC) precipitates of extracellular matrix fluid aspirated from similar cartilages. Most of all of the inhibitory action on an in vitro system of mineral growth shown by whole cartilage PPL-C and by cartilage fluid PPC obtained from noncalcifying sites was contained in the R2 fraction which represented ¼-[unk] of the total hexuronate.

The R2 fraction was diminished or absent from calcified cartilage fluids and from whole calcified epiphyseal septa. The ratio R1 + R2: SR2 ranged from 0.37 to 0.71 in the fluids and whole tissue samples of noncalcified cartilages. The R2 fraction was distinguished from SR2 by a 2- to 3-fold higher protein: hexuronate ratio. These data are interpreted to indicate that the inhibitory R2 fraction was degraded or otherwise inactivated at the zone of provisional calcification and that this inhibitor participates in the physiological mechanism that regulates endochondral calcification.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Campo R. D., Tourtellotte C. D., Bielen R. J. The protein-polysaccharides of articular, epiphyseal plate and costal cartilages. Biochim Biophys Acta. 1969 May 6;177(3):501–511. doi: 10.1016/0304-4165(69)90312-2. [DOI] [PubMed] [Google Scholar]
  2. Campo R. D., Tourtellotte C. D. The composition of bovine cartilage and bone. Biochim Biophys Acta. 1967 Aug 29;141(3):614–624. doi: 10.1016/0304-4165(67)90190-0. [DOI] [PubMed] [Google Scholar]
  3. Di Salvo J., Schubert M. Specific interaction of some cartilage proteinpolysaccharides with freshly precipitating calcium phosphate. J Biol Chem. 1967 Feb 25;242(4):705–710. [PubMed] [Google Scholar]
  4. Hascall V. C., Sajdera S. W. Proteinpolysaccharide complex from bovine nasal cartilage. The function of glycoprotein in the formation of aggregates. J Biol Chem. 1969 May 10;244(9):2384–2396. [PubMed] [Google Scholar]
  5. Hirschman A., Dziewiatkowski D. D. Protein-polysaccharide loss during endochondral ossification: immunochemical evidence. Science. 1966 Oct 21;154(3747):393–395. doi: 10.1126/science.154.3747.393. [DOI] [PubMed] [Google Scholar]
  6. Howell D. S., Carlson L. Alterations in the composition of growth cartilage septa during calcification studied by microscopic x-ray elemental analysis. Exp Cell Res. 1968 Jul;51(1):185–195. doi: 10.1016/0014-4827(68)90169-9. [DOI] [PubMed] [Google Scholar]
  7. Howell D. S., Pita J. C., Marquez J. F., Gatter R. A. Demonstration of macromolecular inhibitors of calcification and nucleational factors in fluid from calcifying sites in cartilage. J Clin Invest. 1969 Apr;48(4):630–641. doi: 10.1172/JCI106021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Howell D. S., Pita J. C., Marquez J. F., Madruga J. E. Partition of calcium, phosphate, and protein in the fluid phase aspirated at calcifying sites in epiphyseal cartilage. J Clin Invest. 1968 May;47(5):1121–1132. doi: 10.1172/JCI105801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Howell D. S., Pita J. C., Marquez J. F. Ultramicro spectrophotometric determination of calcium in biologic fluids. Anal Chem. 1966 Mar;38(3):434–438. doi: 10.1021/ac60235a015. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Pal S., Doganges P. T., Schubert M. The separation of new forms of the proteinpolysaccharides of bovine nasal cartilage. J Biol Chem. 1966 Sep 25;241(18):4261–4266. [PubMed] [Google Scholar]
  12. Pamer T., Glass G. B., Horowitz M. I. Purification and characterization of sulfated glycoproteins and hyaluronidase-resistant mucopolysaccharides from dog gastric mucosa. Biochemistry. 1968 Nov;7(11):3821–3829. doi: 10.1021/bi00851a006. [DOI] [PubMed] [Google Scholar]
  13. Sajdera S. W., Hascall V. C. Proteinpolysaccharide complex from bovine nasal cartilage. A comparison of low and high shear extraction procedures. J Biol Chem. 1969 Jan 10;244(1):77–87. [PubMed] [Google Scholar]
  14. WEINSTEIN G. M., SACHS C. R., SCHUBERT M. PROTEIN POLYSACCHARIDE IN CONNECTIVE TISSUE: INHIBITION OF PHASE SEPARATION. Science. 1963 Nov 22;142(3595):1073–1075. doi: 10.1126/science.142.3595.1073. [DOI] [PubMed] [Google Scholar]

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