High Capacity Calcium-Binding Proteins as Intermediate Calcium Carriers in Biological Mineralization

  • Mary E. Marsh


High-capacity calcium-binding macromolecules have been identified at mineralization fronts in heterodont bivalves, vertebrate tooth dentin and unicellular algae. They appear to be functionally similar to casein, the mineral ion carrier of milk. The bivalve phosphoprotein studied in this laboratory occurs as discrete high molecular weight (MW) particles in the hemolymph and extrapallial fluid and at the inner shell surface during mineral deposition. The phosphoprotein particles are characterized by a high content of aspartic acid, phosphoserine and histidine residues which comprise 80% or more of the peptide chain. The native particles contain a protected pool of calcium and phosphate ions and an exchangeable pool of calcium and magnesium ions. The phosphoprotein monomers are covalently cross-linked via histidinoalanine residues and ionically cross-linked via divalent cations into compact particles with a MW of about 50 million. High concentrations of phosphoprotein particles accumulate at the mineralization front during stimulated biomineralization, but neither inhibit mineral deposition nor influence the crystal habits. The particles are probably calcium ion transporters. However, the mechanism by which protein bound calcium is utilized to form calcium carbonate crystals is unknown.

The bivalve phosphoprotein particles share common characteristics with vertebrate tooth dentin and algal coccolithosomes which have been studied in other laboratories. All three are high-capacity calcium-binding macromolecules which have been localized at mineralization fronts. Both the bivalve phosphoprotein particles and dentin phosphophoryn are aspartic acid-rich, highly phosphosylated proteins which have a tendency to cross-link and fragment. Both coccolithosomes and the bivalve phosphoprotein occur as discrete high MW particles about 20–40 nm in diameter surrounding growing calcium carbonate crystals in vivo. It is postulated that intermediate calcium carriers in the form of high-capacity calcium-binding macromolecules may be a general phenomenon in the biological calcification of skeletal tissues.


Calcium Phosphate Casein Micelle Amorphous Calcium Phosphate Mineralization Front Bovine Dentin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. ADDADI, L., and WEINER, S., 1985. Interactions between acidic proteins and crystals: stereochemical requirements in biomineralization. Proc. Nat. Acad. Sci. U.S.A., 82:4110–4114.CrossRefGoogle Scholar
  2. BUTLER, W.T., BROWN, M., DIMUZIO, M.T., COTHRAN, W.C., and LINDE, A., 1983. Multiple Forms of rat dentin phosphorproteins. Arch. Biochem. Biophys., 225:178–186.PubMedCrossRefGoogle Scholar
  3. CRENSHAW, MA, 1972. The soluble matrix from Mercenaria mercenaria shell. Biomineralization, 6:6–11.Google Scholar
  4. CRENSHAW, MA., and NEFF, J.M. 1969. Decalcification at the mantle-shell interface in mollusks. Amer. Zool., 9:881–885.Google Scholar
  5. DIMUZIO, M.T., and VEIS, A., 1978. The biosynthesis of phosphophoryns and dentin collagen in the continuously erupting rat incisor. J. Biol. Chem., 253:6845–6852.PubMedGoogle Scholar
  6. FUJISAWA, R., KUBOKI, Y., and SASAKI, S., 1985. In vivo cleavage of dentin phosphophoryn following ß-elimination of its phosphoserine residues. Arch. Biochem. Biophys., 243:619–623.PubMedCrossRefGoogle Scholar
  7. HOLT, C., 1982. Inorganic constituents of milk. III. The colloidial calcium phosphate of cow’s milk. J. Dairy Res., 49:29–38.PubMedCrossRefGoogle Scholar
  8. KUBOKI, Y., FUJISAWA, R., AOYAMA, K., and SASAKI, S., 1979. Calcium-specific precipitation of dentin phosphoprotein: a new method of purification and the significance for the mechanism of calcification. J. Den. Res., 58:1926–1932.CrossRefGoogle Scholar
  9. LEE, S.L., KOSSIVA, D., and GLIMCHER, M.J., 1983a. Phosphoproteins of bovine dentin: evidence for polydispersity during tooth maturation. Biochemistry 22:2596–2601.CrossRefGoogle Scholar
  10. LEE, S.L, GLONEK, T., and GLIMCHER, M.J., 1983b. Nuclear magnetic resonance spectroscopic evidence for ternary complex formation of fetal dentin phosphorprotein with calcium and inorganic orthophosphate ions. Calc. Tiss. Intern., 35:815–818.CrossRefGoogle Scholar
  11. LEE, S.L., VEIS, A., and GLONEK, T., 1977. Dentin phosphoprotein: an extracellular calcium-binding protein. Biochemistry, 16:2971–2979.PubMedCrossRefGoogle Scholar
  12. LYSTER, R.L.J., MANN, S., PARKER, S.B., and WILLIAMS, RT.P., 1984. Nature of micellar calcium phosphate in cows’ milk as studied by high-resolution electron microscopy. Biochim. Biophys. Acta, 801:315–317.PubMedCrossRefGoogle Scholar
  13. MARSH, M.E., 1986a. Histidinoalanine, a naturally occurring cross-link derived from phosphoserine and histidine residues in mineral-binding phosphoproteins. Biochemistry, 25:2392–2396.CrossRefGoogle Scholar
  14. MARSH, M.E., 1986b. Biomineralization in the presence of calcium-binding phosphoprotein particles. J. Exp. Zool., 239:207–220.CrossRefGoogle Scholar
  15. MARSH, M.E. and SASS, R.L., 1985. Distribution and characterization of mineral-binding phosphoprotein particles in Bivalvia. J. Exp. Zool., 234:237–242.PubMedCrossRefGoogle Scholar
  16. MARSH, M.S., and SASS, R.L.; 1984. Phosphoprotein particles: calcium and inorganic phosphate binding structures. Biochemistry, 23:1448–1456.PubMedCrossRefGoogle Scholar
  17. MARSH, M.E., and SASS, R.L., 1983. Calcium-binding phosphoprotein particles in the extrapallial fluid and innermost shell lamella of clams. J. Exp. Zool., 266:193–203.CrossRefGoogle Scholar
  18. MASTERS, P.M., 1985. In vivo decomposition of phosphoserine and serine in noncollagenous protein from human dentin. Calc. Tiss. Intern., 37:236–241.CrossRefGoogle Scholar
  19. MCGANN, T.C.A., KEARNEY, RD., BUCHHEIM, W., POSNER, A.S., BETTS, F. and BLUMENTHAL, N.C., 1983. Amorphous calcium phosphate in casein micelles of bovine milk. Calc. Tiss. Intern., 35:821–823.CrossRefGoogle Scholar
  20. MEPHAM, T.B., GAYE, P., and MERCIER, J.C., 1982. Biosynthesis of milk proteins. In Developments in Dairy Chemistry-1 (ed. P. F. Fox), pp. 115–156. New York: Elsevier.Google Scholar
  21. OUTKA, D.E., and WILLIAMS, D.C., 1971. Sequential coccolith morphogenesis in Hymenomonas carterae. J. Protozool., 18:285–297.PubMedGoogle Scholar
  22. PITA, J.C., CUERVO, L.A., MADRUGA, J.E., MULLER, F.J., and HOWELL, D.S., 1970. Evidence for a role of protein polysaccharides in regulation of mineral phase separation in calcifing cartilage. J. Clin. Invest., 49:2188–2197.PubMedCrossRefGoogle Scholar
  23. PRINCE, C.W., OOSAWA, T., BUTLER, W.T., TOMANA, M., BROWN, A.S., BHOWN, M., and SCHROHENLOHER, RE., 1987. Isolation, characterization, and biosynthesis of a phosphorylated glycoprotein from rat bone. J. Biol. Chem., 262:2900–2907.PubMedGoogle Scholar
  24. RUNNEGAR, B., 1989. This volume.Google Scholar
  25. SASS, RL. and MARSH, M.E., 1983. N t-and Np Jr-Histidinoalanine: naturally occurring cross-linking amino acids in calcium-binding phosphoproteins. Biochem. Biophys. Res. Comm., 114:304–309.PubMedCrossRefGoogle Scholar
  26. SCHMIDT, D.G., 1982. Association of caseins and casein micelle structure. In Developments in Dairy Chemistry-1 (ed. P.F. Fox, pp. 61–86). New York, Elsevier.Google Scholar
  27. SCHMIDT, D.G., 1980. Colloidial aspects of casein. Netherlands Milk Dairy J., 34:42–64.Google Scholar
  28. SIKES, C.S., and WHEELER, A.P., 1983. A systematic approach to some fundamental questions of carbonate calcification. In Biomineralization and Biological Metal Ion Accumulation (ed. P. Westbroek and E.W. de Jong), pp. 285–289. Dordrecht: D. Reidel Publishing Co.CrossRefGoogle Scholar
  29. SILER-SIEVENSON, W.G., and VEIS, A., 1983. Bovine dentin phosphophryn: composition and molecular weight. Biochemistry, 22:4326–4335.Google Scholar
  30. SWAISGOOD, H.E., 1982. Chemistry of milk protein. In Developments in Dairy Chemistry-1 (ed. P.F. Fox), pp. 1–59. New York: Elsevier.Google Scholar
  31. VAN DER WAL, P., DE JONG, E.W., WESTBROEK, P., DE BRUIJN, W.C., and MULDERSTAPEL, A.A., 1983a. Polysaccharide localization, coccolith formation, and Golgi dynamics in the Coccolithiphorid Hymenomonas carterae. J. Ultrastr. Res., 85:139–158.Google Scholar
  32. VAN DER WAL, P., DE JONG, E.W., WESTBROEK, P., and DE BRUIJN, W.C., 1983b. Calcification in the Coccolithophorid alga Hymenomonas carterae. In Environmental Biogeochemistry, Ecological Bulletin No. 35 (ed. R Hallberg), pp. 251–258. Stockholm: Publishing House/FRN.Google Scholar
  33. VEIS, DJ., ALBINDER, T.M., CLOHISY, J., RANIMA, M., SABSAY, B., and VEIS, A., 1986. Matrix proteins of the teeth of the sea urchin Lytechinus variegatus. J. Exp. Zool., 240:35–46.PubMedCrossRefGoogle Scholar
  34. WILBUR, KM., and BERNHARDT, A.M., 1982. Mineralization of molluscan shell: effects of free and polyamino acids on crystal growth rate in vitro. Amer. Zool., 22:952.Google Scholar

Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • Mary E. Marsh
    • 1
  1. 1.Dental Science InstituteUniversity of Texas Health Science CenterHoustonUSA

Personalised recommendations