Skip to main content

Advertisement

Log in

Selective adsorption of porcine-amelogenins onto hydroxyapatite and their inhibitory activity on hydroxyapatite growth in supersaturated solutions

  • Laboratory Investigations
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Summary

The selective adsorption of amelogenins onto synthetic hydroxyapatite (HA) and their inhibitory activity on the seeded HA crystal growth were investigated using enamel proteins obtained from the outer layer of immature porcine-enamel (soft, cheeselike in consistency) of developing permanent incisors. Special interests were paid to the effect of a postsecretory degradation of the original amelogenin(s) on their adsorption and inhibitory activity. In the adsorption studies, it was apparent that the originally secreted amelogenin (25 K), proline, and histidine-rich protein (2a), as well as the higher molecular weight components (60–90 K), showed a strong adsorption affinity onto the HA. This adsorption of protein 2a was related to its inhibition of the crystal growth of seeded HA in a dilute supersaturated solution. On the other hand, the partially degraded product (20 K) of amelogenins, protein 2b, lost the high adsorption affinity onto the HA, and consequently showed no significant inhibitory activity. The observed selective adsorption of protein 2a onto HA was apparent at pH 6.0 and pH 7.4 even in the presence of dissociative solvents, such as 3 M urea or 2 and 4 M guanidine-HCl; however, this selective behavior was sensitive to changes in pH, and was not displayed at pH values of 7.8 or 10.8. The results suggest that the originally secreted amelogenin 2a may play an active role in amelogenesis, and that enamel mineralization could be regulated by the secretion of amelogenins and their inactivation through partial enzymic degradation, prior to their complete removal.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Williams RJP (1984) An introduction to biomaterials and the role of organic molecules in their formation. Phil Trans R Soc Lond B 304:411–424

    CAS  Google Scholar 

  2. Termine JD, Belcourt AB, Conn KM, Kleinman HD (1981) Mineral and collagen-binding protein of fetal calf bone. J Biol Chem 256:10403–10408

    CAS  PubMed  Google Scholar 

  3. Romberg RW, Werness PG, Lollar P, Riggs BL, Mann KG (1985) Isolation and characterization of native adult osteonectin. J Biol Chem 260:2728–2736

    CAS  PubMed  Google Scholar 

  4. Robinson C, Lowe NR, Weatherell JA (1977) Changes in aminoacid composition of developing rat incisor enamel. Calcif Tissue Res 23:19–31

    Article  CAS  PubMed  Google Scholar 

  5. Eastoe JE (1979) Enamel protein chemistry— past, present and future. J Dent Res 58(B):753–763

    CAS  PubMed  Google Scholar 

  6. Glimcher MJ (1979) Phosphopeptides of enamel matrix J Dent Res 58(B):790–806

    CAS  PubMed  Google Scholar 

  7. Termine JD, Belcourt AB, Christner PJ, Conn KM, Nylen MV (1980) Properties of dissociatively extracted fetal tooth matrix proteins. I. Principal molecular species in developing bovine enamel. Biol Chem 255:9760–9768

    CAS  Google Scholar 

  8. Robinson C, Kirkham J (1984) Enamel matrix components. Alterations during development and possible interactions with the mineral phase. In: Fearnhead RW, Suga S (ed) Tooth enamel IV, Elsevier, Amsterdam, pp 261–265

    Google Scholar 

  9. Doi Y, Eanes ED, Shimokawa H, Termine JD (1984) Inhibition of seed growth of enamel apatite crystals by amelogenin and enamelin protein in vitro. J Dent Res 63:98–105

    CAS  PubMed  Google Scholar 

  10. Bai P, Warshawsky H (1985) Morphological studies on the distribution of enamel matrix proteins using routine electron microscopy and freeze-fracture replicas in the rat incisor Anat Rec 212:1–16

    Article  CAS  PubMed  Google Scholar 

  11. Kallenbach E (1982) Fine structure of extracted rat incisor enamel. J Dent Res 61 (Special Issue):1515–1523

    Google Scholar 

  12. Jodaikin A, Traub W, Weiner S (1986) Protein conformation in rat tooth enamel. Archs Oral Biol 31:685–689

    Article  CAS  Google Scholar 

  13. Hayashi Y, Bianco P, Shimokawa H, Termine JD, Bonucci E (1986) Organic-inorganic relationships, and immunohistochemical localization of amelogenins and enamelins in developing enamel. Bas Appl Histochem 30:291–299

    CAS  Google Scholar 

  14. Zeichner-David M, Macdougall M, Vides J, Snead M, Slavkin HC (1984) Enamel gene expression in selected vertebrates. In: Fearnhead RW, Suga S (ed) Tooth enamel IV. Elsevier, Amsterdam, pp 183–187

    Google Scholar 

  15. Graham EE (1985) Isolation of enamelinelike proteins from blue shark (Prionace Glauca) enameloid J Exp Zool 234:185–191

    Article  CAS  PubMed  Google Scholar 

  16. Snead ML, Lau EC, Zeichner-David M, Fincham AG, Woo SLC, Slavkin HC (1985) DNA sequence for cloned cDNA murine amelogenin reveal the amino acid sequence for enamel-specific protein. Biochem Biophys Res Comm 129:812–818

    Article  CAS  PubMed  Google Scholar 

  17. Takagi T, Suzuki M, Baba T, Minegishi K, Sasaki S (1984) Complete amino acid sequence of amelogenin in developing bovine enamel. Biochem Biophys Res Comm 121:592–597

    Article  CAS  PubMed  Google Scholar 

  18. Fukae M, Shimizu M (1985) Amino acid sequence of a protein from developing porcine enamel. Jpn J Oral Biol 25 (Suppl):29

    Google Scholar 

  19. Seyer JM, Glimcher MJ (1977) Evidence for the presence of numerous protein components in immature bovine dental enamel. Calcif Tissue Res 24:253–257

    Article  CAS  PubMed  Google Scholar 

  20. Shimokawa H, Sasaki S (1978) Study on the biosynthesis of bovine enamel protein in vitro. J Dent Res 57:133–138

    CAS  PubMed  Google Scholar 

  21. Robinson C, Briggs HD, Atkinson PJ, Weatherell JA (1979) Matrix and mineral changes in developing enamel. J Dent Res 58 (Special Issue B):871–880

    CAS  PubMed  Google Scholar 

  22. Fukae M, Tanabe T, Ijiri H, Shimizu M (1980) Studies on porcine enamel protein: a possible original enamel protein. Tsurumi University Dent J 6:87–94

    CAS  Google Scholar 

  23. Fincham AG, Belcourt AB, Termine JD, Butler WT, Cothran WC (1983) Amelogenins: sequence homologies in enamel-matrix proteins from three manmalian species. Biochem J 211:149–154

    CAS  PubMed  Google Scholar 

  24. Sasaki S, Shimokawa H, Tanaka K (1982) Biosynthesis of rat enamel matrix components in vivo J Dent Res 61 (Special Issue):1479–1482

    CAS  Google Scholar 

  25. Strawich E, Glimcher MJ (1985) Synthesis and degradation in vivo of a phosphoprotein from rat dental enamel. Identification of a phosphorylated precursor protein in the extracellular organic matrix. Biochem J 230:423–433

    CAS  PubMed  Google Scholar 

  26. Fukae M, Ijiri H, Tanabe T, Shimizu M (1979) Partial amino acid sequences of two proteins in developing porcine enamel. J Dent Res 58:1000–1001

    CAS  PubMed  Google Scholar 

  27. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  28. Aoba T, Moreno EC (1984) Hydroxyapatite preparation and crystal growth on hydroxyapatite seeds. J Dent Res 63:874–880

    CAS  PubMed  Google Scholar 

  29. Shimizu M, Fukae M (1983) Enamel proteins. In: Suga S (ed) Mechanisms of tooth enamel formation. Quintessence Publishing Co., Tokyo-Berlin-Chicago-Rio de Janeiro, pp 125–141

    Google Scholar 

  30. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  31. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Chem 72:248–254

    CAS  Google Scholar 

  32. Moreno EC, Kresak M, Zaradnik RT (1977) Physicochemical aspects of fluoride-apatite systems relevant to the study of dental caries. Caries Res (Suppl 1) 11:142–171

    Article  PubMed  Google Scholar 

  33. Vogel AJ (1961) Quantitative inorganic analysis, 3rd ed. John Wiley & Sons, New York, p 810

    Google Scholar 

  34. Yanagisawa T, Nylen MU (1980) The relationship between matrix and mineral in rat enamel—an electron microscope study. J Dent Res 59:343

    Google Scholar 

  35. Moreno EC, Varughese K, Hay DI (1979) Effect of human salivary proteins on the precipitation kinetics of calcium phosphate. Calcif Tissue Int 28:7–16

    Article  CAS  PubMed  Google Scholar 

  36. Aoba T, Moreno EC, Hay DI (1984) Inhibition of apatite crystal growth by the amino-terminal segment of human salivary acidic proline-rich proteins. Calcif Tissue Int 36:651–658

    Article  CAS  PubMed  Google Scholar 

  37. Shimizu M, Tanabe T, Fukae M (1979) Proteolytic enzyme in porcine immature enamel. J Dent Res 58(b):782–789

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aoba, T., Fukae, M., Tanabe, T. et al. Selective adsorption of porcine-amelogenins onto hydroxyapatite and their inhibitory activity on hydroxyapatite growth in supersaturated solutions. Calcif Tissue Int 41, 281–289 (1987). https://doi.org/10.1007/BF02555230

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02555230

Key words

Navigation