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Mineral induction by immobilized polyanionic proteins

Calcified Tissue International volume 44, pages 286–295 (1989)Cite this article

Summary

The purpose of this study was to investigate the mineral induction capacityin vitro of polyanionic proteins covalently bound to a surface. Rat dentin γ-carboxyglutamate-containing protein of the osteocalcin type (Gla-protein), proteoglycan (PG), and phosphoprotein (PP-H), as well as phosvitin (PhV) and bovine serum albumin (BSA), were covalently linked to agarose beads. These were incubated at 37°C in solutions with a Ca/P molar ratio of 1.67, [Ca][P] molar products in the range 1.0–1.8 mM2, and an ionic strength of 0.165. The incubations were performed at constant pH and composition conditions; no spontaneous precipitation occurred under these conditions. Mineral formation, as monitored by scanning electron microscopy (SEM), was induced by all immobilized polyanions, including enzymatically dephosphorylated PP-H and PhV. No mineral was induced by BSA. The mineral inductive capacity of immobilized polyanionic proteins, as judged by the SEM after identical incubations, was found to differ between the different ligands. The mineral induced by PP-H and PG was shown by X-ray diffraction to be apatitic. It was concluded that, although polyanionic proteins in solution may inhibit mineral induction and growth, very minute quantities of such molecules, when immobilized on a surface, induce mineral at physiological concentrations of calcium and phosphate ions. The data presented may be taken to suggest that PP-H and PG, and perhaps other polyanions, may possibly be responsible for mineral nucleation in dentin and bone. The results, however, also point to the rather limited specificity in this type of reaction.

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References

  1. Boskey AL (1985) Overview of cellular elements and macromolecules implicated in the initiation of mineralization. In: Butler WT (ed) The chemistry and biology of mineralized tissues. EBSCO Media, Birmingham, AL, pp 335–343

    Google Scholar 

  2. Linde A (1984) Noncollagenous proteins and proteoglycans in dentinogenesis. In: Linde A (ed) Dentin and dentinogenesis. CRC Press, Boca Raton, FL, pp 55–92

    Google Scholar 

  3. Linde A (1985) Dynamic aspects of dentinogenesis. In: Butler WT (ed) The chemistry and biology of mineralized tissues. Ebsco Media, Birmingham, AL, pp 344–355

    Google Scholar 

  4. Veis A (1985) Phosphoproteins of dentin and bone. Do they have a role in matrix mineralization? In: Butler WT (ed) The chemistry and biology of mineralized tissues. EBSCO Media, Birmingham, AL, pp 170–176

    Google Scholar 

  5. Landis WJ, Paine MC, Glimcher MJ (1977) Electron microscopic observations of bone tissue prepared anhydrously in organic solvents. J Ultrastruct Res 59:1–30

    PubMed  Article  CAS  Google Scholar 

  6. Williams RJP (1977) Calcium chemistry and its relation to protein binding. In: Wasserman RH, Corradino RA, Carafoli E, Kretsinger RH, MacLennan DH, Siegel FL (eds) Calcium-binding proteins and calcium function. Elsevier/North-Holland, New York, pp 3–12

    Google Scholar 

  7. Zanetti M, de Bernard B, Jontell M, Linde A (1981) Ca2+-binding studies of the phosphoprotein from rat-incisor dentine. Eur J Biochem 113:541–545

    PubMed  Article  CAS  Google Scholar 

  8. Cookson DJ, Levine BA, Williams RJP, Jontell M, Linde A, de Bernard B (1980) Cation binding by the rat-incisor-dentine phosphoprotein. A spectroscopic investigation. Eur J Biochem 110:273–278

    PubMed  Article  CAS  Google Scholar 

  9. Stetler-Stevenson WG, Veis A (1987) Bovine dentin phosphophoryn: calcium ion binding properties of a high molecular weight preparation. Calcif Tissue Int 40:97–102

    PubMed  CAS  Google Scholar 

  10. Linde A, Hansson H-A (1983) Localization of Gla-proteins during calcification. In: de Bernard B, Sottocasa GL, Sandri G, Caraafoli E, Taylor AN, Vanaman TC, Williams RJP (eds) Calcium-binding proteins 1983. Elsevier, Amsterdam, pp 65–66

    Google Scholar 

  11. Jontell M, Linde A (1983) Non-collagenous proteins of predentine from dentinogenically active bovine teeth. Biochem J 214:769–776

    PubMed  CAS  Google Scholar 

  12. Weinstock M, Leblond CP (1973) Radioautographic visualization of the deposition of a phosphoprotein at the mineralization front in the dentin of the rat incisor. J Cell Biol 56:838–845

    PubMed  Article  CAS  Google Scholar 

  13. MacDougall M, Zeichner-David M, Slavkin HC (1985) Production and characterization of antibodies against murine dentin phosphoprotein. Biochem J 232:493–500

    PubMed  CAS  Google Scholar 

  14. Sundström B (1971) New aspects on the utilization of inorganic sulphate during dentin formation. Histochemie 26:61–66

    PubMed  Article  Google Scholar 

  15. Howell DS, Pita JC (1976) Calcification of growth plate cartilage with special reference to studies on micropuncture fluid. Clin Orthop 118:208–229

    PubMed  CAS  Google Scholar 

  16. Blumenthal NC, Posner AS, Silverman LD, Rosenberg LC (1979) Effect of proteoglycans on in vitro hydroxyapatite formation. Calcif Tissue Int 27:75–82

    PubMed  Article  CAS  Google Scholar 

  17. Chen C-C, Boskey AL, Rosenberg LC (1984) The inhibitory effect of cartilage proteoglycan on hydroxyapatite growth. Calcif Tissue Int 36:285–290

    PubMed  Article  CAS  Google Scholar 

  18. Romberg RW, Werness PG, Riggs BL, Mann KG (1986) Inhibition of hydroxyapatite crystal growth by bone-specific and other calcium-binding proteins. Biochemistry 25:1176–1180

    PubMed  Article  CAS  Google Scholar 

  19. Nawrot CF, Campbell DJ, Schroeder JK, van Valkenburg M (1976) Dental phosphoprotein-induced formation of hydroxyapatite during in vitro synthesis of amorphous calcium phosphate. Biochemistry 15:3445–3449

    PubMed  Article  CAS  Google Scholar 

  20. Termine JD, Conn KM (1976) Inhibition of apatite formation by phosphorylated metabolites and macromolecules. Calcif Tissue Res 22:149–157

    PubMed  Article  CAS  Google Scholar 

  21. Termine JD, Eanes ED, Conn KM (1980) Phosphoprotein modulation of apatite crystallization. Calcif Tissue Int 31:247–251

    PubMed  Article  CAS  Google Scholar 

  22. Udich H-J, Höft HD, Börnig H (1986) Effect of phosphoprotein on precipitation and crystallization of calcium phosphate salts. An in vitro study using an agar gel matrix model. Biomed Biochim Acta 6:703–711

    Google Scholar 

  23. Linde A, Bhown M, Butler WT (1980) Noncollagenous proteins of dentin. J Biol Chem 255:5931–5942

    PubMed  CAS  Google Scholar 

  24. Linde A, Bhown M, Cothran WC, Höglund A, Butler WT (1982) Evidence for several γ-carboxyglutamate acid-containing proteins in dentin. Biochim Biophys Acta 704:235–239

    PubMed  CAS  Google Scholar 

  25. Linde A, Jontell M, Lundgren T, Nilson B, Svanberg U (1983) Noncollagenous proteins of rat compact bone. J Biol Chem 258:1698–1705

    PubMed  CAS  Google Scholar 

  26. Fleisch H, Neuman WF (1961) Mechanisms of calcification: role of collagen, polyphosphates and phosphatase. Am J Physiol 200:1296–1300

    CAS  Google Scholar 

  27. Crenshaw MA, Ramp WK, Gonnerman WA, Toverud SU (1974) Effects of dietary vitamin D levels on the in vitro mineralization of chick metaphyses. Proc Soc Exp Biol Med 146:488–493

    PubMed  CAS  Google Scholar 

  28. Blumenthal NC, Posner AS, Holmes JM (1972) Effect of preparation on the properties and transformation of amorphous calcium phosphate. Mat Res Bull 7:1181–1190

    Article  CAS  Google Scholar 

  29. Taves DR, Neuman WF (1964) Factors controlling calcification in vitro: fluoride and magnesium. Arch Biochem Biophys 108:390–397

    PubMed  Article  CAS  Google Scholar 

  30. Termine JD, Eanes ED (1974) Calcium phosphate deposition from balanced salt solutions. Calcif Tissue Res 15:81–84

    PubMed  Article  CAS  Google Scholar 

  31. Chen PS, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756–1758

    Article  CAS  Google Scholar 

  32. Neuman WF, Neuman MW (1958) The chemical dynamics of bone mineral, University of Chicago Press, Chicago, IL

    Google Scholar 

  33. Nancollas GH (1976) The kinetics of crystal growth and renal stone-formation. In: Fleisch H, Robertson WG, Smith LH, Vahlensieck W (eds) Urolithiasis research. Plenum Press, New York, pp 5–23

    Google Scholar 

  34. Howell DS, Pita JC, Marquez JF (1968) Partition of calcium, phosphate and protein in the fluid aspirated at calcifying sites in epiphyseal cartilage. J Clin Invest 47:1121–1131

    PubMed  CAS  Google Scholar 

  35. Nancollas GH, Tomazic B (1974) Growth of calcium phosphate on hydroxyapatite crystals. Effect of supersaturation and ionic medium. J Phys Chem 78:2218–2225

    Article  CAS  Google Scholar 

  36. Koutsoukos PG, Nancollas GH (1981) Crystal growth of calcium phosphates—epitaxial considerations. J Cryst Growth 53:10–19

    Article  CAS  Google Scholar 

  37. Davies CW (1962) Ion association. Butterworths, London

    Google Scholar 

  38. Plummer LN, Jones BF, Truesdale AH (1984) WATEQF—A Fortran IV version of WATEQ, a computer program for calculating chemical equilibrium of natural waters U.S. Geological Survey, Reston

    Google Scholar 

  39. Boskey AL, Posner AS (1976) Formation of hydroxyapatite at low supersaturation. J Phys Chem 80:40–45

    Article  CAS  Google Scholar 

  40. Taborsky G (1983) Phosvitin. Adv Inorg Biochem 5:235–279

    PubMed  CAS  Google Scholar 

  41. Termine JD, Kleinman HK, Whitson SW, Conn KM, McGarvey ML, Martin GR (1981) Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26:99–105

    PubMed  Article  CAS  Google Scholar 

  42. Franzén A, Heineg»rd D (1985) Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochem J 232:715–724

    PubMed  Google Scholar 

  43. Heineg»rd D, Paulsson M (1984) Structure and metabolism of proteoglycans. In: Piez KA, Reddi AH (eds) Extracellular matrix biochemistry. Elsevier, New York, pp 277–328

    Google Scholar 

  44. Hauschka PV, Gallop PM (1977) Purification and calciumbinding properties of osteocalcin, the γ-carboxy-glutamate-containing protein of bone. In: Wasserman RH, Corradino RA, Carafoli E, Kretsinger RH, Mac-Lennan DH, Siegel FL (eds) Calcium-binding proteins and calcium function. Elsevier/North Holland, Amsterdam, pp 338–347

    Google Scholar 

  45. Price PA, Otsuka AS, Poser JW (1977) Comparison of γ-carboxyglutamic acid-containing proteins from bovine and swordfish bone: primary structure and Ca2+ binding. In: Wasserman RH, Corradino RA, Carafoli E, Kretsinger RH, MacLennan DH, Siegel FL (eds) Calcium-binding proteins and calcium function. Elsevier/North Holland, New York. pp 333–337

    Google Scholar 

  46. Svärd M, Drakenberg T, Andersson T, Fernlund P (1986) Calcium binding to bone γ-carboxyglutamic acid protein from calf studied by43Ca NMR. Eur J Biochem 158:373–378

    PubMed  Article  Google Scholar 

  47. Grizzuti K, Perlmann GE (1973) Binding of magnesium and calcium ions to the phosphoglycoprotein phosvitin. Biochemistry 12:4399–4403

    PubMed  Article  CAS  Google Scholar 

  48. Butler WT (1984) Dentin collagen: chemical structure and role in mineralization. In: Linde A (ed) Dentin and dentinogenesis. CRC Press, Boca Raton, pp 37–53

    Google Scholar 

  49. Linde A, Bhown M, Butler WT (1981) Non-collagenous proteins of rat dentin. Evidence that phosphoprotein is not covalently bound to collagen. Biochim Biophys Acta 667:341–350

    PubMed  CAS  Google Scholar 

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Author notes

  1. Anders Linde

    Present address: Department of Oral Biochemistry, Faculty of Odontology, Gothenburg University, S-400 33, Gothenburg, Sweden

  2. Adrian Lussi

    Present address: Department of Operative, Preventive, and Pediatric Dentistry, School of Dentistry, University of Bern, 3010, Bern, Switzerland

Affiliations

  1. Dental Research Center and Departments of Endodontics and Pediatric Dentistry, University of North Carolina, Chapel Hill, North Carolina, USA

    Anders Linde, Adrian Lussi & Miles A. Crenshaw

Authors
  1. Anders Linde
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  2. Adrian Lussi
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  3. Miles A. Crenshaw
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Linde, A., Lussi, A. & Crenshaw, M.A. Mineral induction by immobilized polyanionic proteins. Calcif Tissue Int 44, 286–295 (1989). https://doi.org/10.1007/BF02553763

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Key words

  • Mineralization
  • Polyanions
  • Glaprotein (osteocalcin)
  • Proteoglycan
  • Phosphoprotein