Abstract
Correlations between structural properties of bivalve shell organic matrix and its proposed functions in the regulation of biomineralization were examined using proteinaceous fractions obtained from the shell of the oyster Crassostrea virginica (Gmelin) following dissolution of the mineral with ethylenediaminetetraacetate (EDTA). Matrix isolated in this way contains a continuous size distribution of proteins ranging from relatively small molecular weight (Mr) soluble matrix (SM) components to insoluble matrix (IM) components. Regulation of mineralization by these components was determined primarily by their reduction of crystal growth rate in an in vitro assay. For all fractions tested, there was an inverse correlation between Mr and regulatory activity. However, matrix properties other than molecular size may be important in regulation of crystal growth in vivo in that the larger but less acidic of two soluble matrix proteins was a more effective inhibitor of spicule formation by sea urchin embryos. Base treatment of IM and high molecular weight SM fractions that had little or no inhibitory activity when untreated, resulted in constituents that had a molecular weight distribution and in vitro inhibitory activity in the same range as whole SM. These results, combined with the finding that a major fraction of IM has an amino acid composition very similar to the highly charged SM fractions, suggest that much of the matrix is made up of similar molecules, and that their function in crystal growth regulation may change as they interact to form units of increasing size. A second class of IM was isolated which contained dihydroxyphenylalanine (dopa) and a preponderance of hydrophobic amino acids. This material may represent the basic structural framework of the matrix.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature cited
Addadi, L., Weiner, S. (1985). Interactions between acidic proteins and crystals: stereo-chemical requirements in biomineralization. Proc. natl. Acad. Sci., USA 82: 4110–4114
Anonymous (1973). Dequest 2010, phosphonate for scale and corrosion control, chelation, dispersion. Technical Bulletin No. IC ISCS-317, Mosanto Chemical Company
Anonymous (1983). Acrysol scale inhibitors. Technical Bulletin CS-513, Rohm and Haas Company
Beedham, G. E. (1958). Observations on the non-calcareous component of the shell of the Lamellibranchia. Quart. J. microsc. Sci. 99: 341–357
Bernhardt, A. M., Manyak, D. M., Wilbur, K. M. (1985). In vitro recalcification of organic matrix of scallop shell and serpulid tubes. J. mollusc. Stud. 51: 284–289
Borman, A. H., de Jong, E. W., Huizinga, M., Kok, D. S., Westbroek, P., Bosch, L. (1982). The role in CaCO3 crystallization of an acid Ca2+-binding polysaccharide associated with coccoliths of Emiliania huxley. Eur. J. Biochem. 129: 179–183
Crenshaw, M. A. (1972). The soluble matrix from Mercenaria mercenaria shell. Biomineralization 6: 6–11
Crenshaw, M. A. (1982). Mechanisms of normal biological mineralization of calcium carbonate. In: Nancollas, G. H. (ed.) Biological mineralization and demineralization. Springer-Verlag, Berlin, p. 243–251
Crenshaw, M. A., Ristedt, H. (1976). The histochemical localization of reactive groups in septal nacre from Nautilus pompilius L. In: Watabe, N., Wilbur, K. M. (eds.) The mechanisms of mineralization in the invertebrates and plants. University of South Carolina Press, Columbia, South Carolina, p. 355–367
Davis, N. R., Cavanagh, J. C. (1981). Hard tissue mineralization inhibitors. In: Veis, A. (ed.) The chemistry and biology of mineralized connective tissues. Elsevier-North Holland, New York, p. 489–491
Degens, E. T., Spencer, D. W., Parker, R. H. (1967). Paleobiochemistry of molluscan shell proteins. Comp. Biochem. Physiol. 20: 553–579
Galstoff, P. S. (1964). The American oyster Crassostrea virginica (Gmelin). Fish. Bull. US. 64: US Fish and Wildlife Service
Gordon, J., Carriker, M. R. (1980). Sclerotized protein in the shell matrix of a bivalve mollusc. Mar. Biol. 57: 251–260
Greenfield, E. M., Wilson, D. C., Crenshaw, M. A. (1984). Ionotropic nucleation of calcium carbonate by molluscan matrix. Amer. Zool. 24: 925–932
Grégoire, C. (1972). Structure of the molluscan shell. In: Florkin, M., Sheer, B. (eds.) vol. 7. Academic Press, New York, p. 45–102
Grégoire, C., Duchâteau, G., Florkin, M. (1955). La trame protidique des nacres et des perles. Ann. Inst. Oceanogr., Paris 31: 1–36
Hidaka, H., Tanaka, T. (1983). Naphthalene sulfonamides as calmodulin antagonists. In: Colowick, S. P., Kaplan, M. O. (eds.) Methods in enzymology. vol. 102. pp 185–194. Academic Press, New York, p. 185–194
Kleinbaum, D., Kupper L. (1978). Applied regression analysis and other multivariable methods. Wadsworth, Belmont, California
Krampitz, G., Drolshagen, H., Häusle, T., Hof-Irmscher, K. (1983). Organic matrix of mollusc shells. In: Westbroek, P., de Jong, E. W. (eds.) Biomineralization and biological metal accumulation. D. Reidel Publ. Co., Dordrecht, Holland, p. 231–247
Krampitz, G., Engels, J., Cazaux, C. (1976). Biochemical studies on water-soluble proteins and related components of gastropod shells. In: Watabe, N., Wilbur, K. M. (eds.) The mechanisms of mineralization in the intervebrates and plants. Univ. of South Carolina Press, Columbia, South Carolina, p. 155–173
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. (1951). Protein determination with the Folin phenol reagent. J. Biol. Chem. 193: 265–275
Mann, S. (1983). Mineralization in biological systems. In: Clarke, M. T., Goodenough, J. B., Ibers, J. A., Jorgenson, C. K., Neilands, J. B., Reinen, D., Weiss, R., Williams, R. J. P. (eds.) Structure and bonding, vol. 54. Springer-Verlag, Berlin, p. 125–175
Meenakshi, V. R., Hare, P. E., Wilbur, K. M. (1971). Amino acids of the organic matrix of the gastropod shells. Comp. Biochem. Physiol. 40B: 1037–1043
Menanteau, J., Neuman, W. F., Neumann, W. W. (1982). A study of bone proteins which can prevent hydroxyapatite formation. Metab. Bone Dis. Rel. Res. 4: 157–162
Miller, G. L. (1959): Protein determination for large numbers of samples. Anal. Chem. 31: 1964
Reddy, M. M., Nancollas G. H. (1973). Calcite crystal growth inhibition by phosphonates. Desalination 12: 61–73
Roer, R. D., Dillaman, R. M. (1984). The structure and calcification of the crustacean cuticle. Amer. Zool. 24: 893–910
Sikes, C. S., Okazaki, K., Fink, R. D. (1981). Respiratory CO2 and the supply of inorganic carbon for calcification of sea urchin embryos. Comp. Biochem. Physiol. 70A: 285–291
Sikes, C. S., Wheeler, A. P. (1982). Carbonic anhydrase and carbon dioxide fixation in coccolithophorids. J. Phycol. 18, 423–426
Sikes, C. S., Wheeler, A. P. (1983). A systematic approach to some fundamental questions of carbonate calcification. In: Westbroek, P., de Jong, E. W. (eds.) Biomineralization and biological metal accumulation. D. Reidel Publ. Co., Dordrecht, Holland, p. 285–289
Sikes, C. S., Wheeler, A. P. (1985).Inhibition of inorganic or biological CaCO3 deposition by polyamino acid derivatives. US Patent 4, 534, 881
Sikes, C. S., Wheeler, A. P. (1986). The organic matrix from oyster shell as a regulator of calcification in vivo. Biol. Bull. 170: 494–505
Sikes, C. S., Wheeler, A. P. (1988). Biopolymers from biominerals as regulators of mineralization. CHEMTECH, (In press)
Simkiss, K. (1965). The organic matrix of the oyster shell. Comp. Biochem. Physiol. 16: 427–435
Termine, J. D., Eanes, E. D., Conn, K. M. (1980). Phosphoprotein modulation of apatite crystallization. Calcif. Tissue Int. 31: 247–251
Termine, J. D., Kleinman, H. K., Whitson, S. W., Conn, K. M., McGarvey, M. L., Martin, G. R. (1981). Osteonectin, a bonespecific protein linking mineral to collagen. Cell 36: 99–105
Travis, D. E., Francois, C. J., Bonar, L. C., Glimcher, M. J. (1967). Comparative studies of the organic matrices of invertebrate mineralized tissues. J. Ultrastruct. Res. 18: 519–550
Waite, J. H. (1983). Quinone-tanned soleroproteins. In: Wilbur, K. M. (ed.) The mollusca. vol. 1. Academic Press, New York, p. 467–504
Waite, J. H., Benedict, C. V. (1984). Assay of dihydroxyphenylalanine (dopa) in invertebrate structural proteins. In: Colowick, J. P., Kaplan, N. O. (eds.) Methods in enzymology. vol. 107. Academic Press, New York, p. 297–413
Wallace, R., Tallant, E., Cheung, W. (1980). Assay of calmodulin by Ca2+-dependent phosphodiesterase. In: Cheung, W. (ed.) Calcium and cell function. vol. 1. Academic Press, New York, p. 13–40
Weiner, S. (1979). Aspartic acid-rich proteins: major components of the soluble organic matrix of mollusk shells. Calcif. Tissue Int. 29: 163–167
Weiner, S. (1982). Separation of acidic proteins from mineralized tissues by reversed phase high-performance liquid chromatography. J. Chromatog. 245: 148–154
Weiner, S. (1983). Mollusc shell formation: isolation of two organic matrix proteins associated with calcite deposition in the bivalve Mytilus californianus. Biochemistry 22: 4139–4145
Weiner, S. (1984). Organization of organic matrix components in mineralized tissues. Amer. Zool. 24: 945–951
Weiner, S. (1986): Organization of extracellularly mineralized tissues: A comparative study of biological crystal growth. Crit. Rev. Biochem. 20: 365–408
Weiner, S., Hood, L. (1975). Soluble protein of the organic matrix of mollusc shells: a potential template for shell. Science, N.Y. 190: 987–989
Weiner, S., Traub, W. (1980). X-ray diffraction study of the insoluble organic matrix of mollusk shells. FEBS Lett. 111: 311–316
Weiner, S., Traub, W., Lowenstam, H. A. (1983). Organic matrix in calcified exoskeletons. In: Westbroek, P., de Jong, E. W. (eds.) Biomineralization and biological metal accumulation. D. Reidel Publ. Co., Dordrecht, Holland, p. 205–224
Wheeler, A. P., George, J. W., Evans, C. A. (1981). Control of calcium carbonate crystal nucleation and crystal growth by soluble matrix of oyster shell. Science, N.Y. 212: 1397–1398
Wheeler, A. P., Rusenko, K. W., George, J. W., Sikes, C. S. (1987). Evaluation of calcium binding by oyster soluble matrix and its role in biomineralization. Comp. Biochem. Physiol. 87B: 953–960
Wheeler, A. P., Sikes, C. S. (1984). Regulation of carbonate calcification by organic matrix. Amer. Zool. 24: 933–944
Wheeler, A. P., Sikes, C. S. (1986). Inhibition of the formation of inorganic or biological CaCO3-containing deposits by a proteinaceous fraction obtained from CaCO3-forming organisms. US Patent 4, 587, 021
Whitaker, J. R. (1980). Changes occurring in proteins in alkaline solution. In: Whitaker, J. R., Fugimaki, M. (eds.) Chemical deterioration of proteins. American Chemical Society, Washington, p. 145–163
Wilbur, K. M., Manyak, D. M. (1984). Biochemical aspects of molluscan mineralization. In: Costlow, J. C., Tipper, R. C. (eds.) Marine biodeterioration: an interdisciplinary study. Naval Institute Press, Annapolis, Maryland, p. 30–37
Wilbur, K. M., Watabe, N. (1963). Experimental studies on calcification in molluscs and the alga Coccolithus huxleyi. Ann. N.Y. Acad. Sci. 109: 82–112
Zar, J. H. (1974). Biostatistical analysis. Prentice-Hall, Englewood Cliffs, New Jersey
Author information
Authors and Affiliations
Additional information
Communicated by J. P. Grassle, Woods Hole
Rights and permissions
About this article
Cite this article
Wheeler, A.P., Rusenko, K.W., Swift, D.M. et al. Regulation of in vitro and in vivo CaCO3 crystallization by fractions of oyster shell organic matrix. Marine Biology 98, 71–80 (1988). https://doi.org/10.1007/BF00392660
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392660