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A comparison of Ca2+-, Mg2+-ATPase and alkaline phosphatase activities of rat incisor pulp

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Summary

A plasma membrane preparation derived from incisor pulps of 250-g rats was used as the source of alkaline phosphatase and an ATPase activated by either Ca2+ or Mg2+. Properties of the two enzymes were then compared under a variety of experimental conditions to determine if ATPase activity is clearly distinct from alkaline phosphatase activity. (a) The optimum pH for ATP hydrolysis was 8.0, compared with 10.2 forp-nitrophenylphosphate. (b) At the optimum pH, ATP hydrolysis required either Ca2+ or Mg2+ for activation, whereasp-nitrophenylphosphate hydrolysis did not require and was little influenced by the presence of divalent cations. (c) Alkaline phosphatase showed maximal activity at 40°C and ATPase at 50°C, with complete inactivation at 60°C and 70°C, respectively. (d) When the plasma membrane preparation was preincubated in the absence of substrate at varying temperatures of pH, alkaline phosphatase was less stable than ATPase at extreme pH and high temperatures. (e) Alkaline phosphatase activity was lost more rapidly than ATPase activity during storage at 4°C for up to 10 days. (f) Butanol extraction of the plasma membrane preparation to remove phospholipid destroyed ATPase activity and enhanced alkaline phosphatase activity approximately fivefold. On the basis of these comparisons it is concluded that ATP hydrolysis andp-nitrophenyl-phosphate hydrolysis are the result of separate enzyme activities.

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References

  1. Robison, R.: The possible significance of hexosephosphoric esters in ossification. Biochem. J.17, 286–293 (1923)

    Google Scholar 

  2. Martin, K.L., Melancon, M.J., DeLuca, H.F.: Vitamin D stimulated calcium-dependent adenosine triphosphatase from brush borders of rat small intestine. Biochim. Biophys. Res. Commun.35, 819–823 (1969)

    Google Scholar 

  3. MacLennan, D.H.: Purification and properties of an adenosine triphosphatase from sarcoplasmic reticulum. J. Biol. Chem.245, 4508–4518 (1970)

    PubMed  Google Scholar 

  4. Schatzmann, H.J., Rossi, G.L.: (Ca2++Mg2+)-activated membrane ATPase in human red cells and their possible relations to cation transport. Biochim. Biophys. Acta241, 379–392 (1971)

    PubMed  Google Scholar 

  5. Ali, S.Y., Sajdera, S.W., Anderson, H.C.: Isolation and characterization of calcifying matrix vesicles from epiphyseal cartilage. Proc. Natl. Acad. Sci. U.S.A.67, 1513–1520 (1970)

    PubMed  Google Scholar 

  6. Heyden, G., From, S.H.: A histochemical demonstration of ATPase activity during tooth ontogeny in the mouse. Arch. Oral Biol.14, 1125–1232 (1969)

    PubMed  Google Scholar 

  7. Severson, A.R.: Histochemical demonstration of nucleoside triphosphate hydrolysis in the mouse dentition. Acta Histochem.40, 86–97 (1971

    PubMed  Google Scholar 

  8. Guo, M.K., Messer, H.H.: Properties of Ca2+-, Mg2+-activated adenosine triphosphatase from rat incisor pulp. Arch. Oral Biol.21, 637–640 (1976)

    PubMed  Google Scholar 

  9. Goggins, J.F., Fullmer, H.M.: Hydrolytic enzyme histochemistry of the rat molar pulp. Arch. Oral Biol.12, 639–644 (1967)

    PubMed  Google Scholar 

  10. Haussler, M.R., Nagode, L.A., Rasmussen, H.: Induction of intestinal brush border alkaline phosphatase by vitamin D, identity with Ca-ATPase Nature228, 1199–1201 (1970)

    PubMed  Google Scholar 

  11. Russell, R.G.G., Monod, A., Bonjour, J.P., Fleisch, H.: Relation between alkaline phosphatase and Ca2+-ATPase in calcium transport. Nature [New Biol.]240, 126–127 (1972)

    Google Scholar 

  12. Ramp, W.K.: Cellular control of calcium movement in bone: Interrelationships of the bone membrane, parathyroid hormone and alkaline phosphatase. Clin. Orthop. Rel. Res.106, 311–322 (1975)

    Google Scholar 

  13. Felix, R., Fleisch, H.: Pyrophosphatase and ATPase of isolated cartilage matrix vesicles. Calcif. Tissue Res.22, 1–7 (1976)

    PubMed  Google Scholar 

  14. Messer, H.H., Rogers, J., Shami, Y., Copp, D.H.: Ca2+-, Mg2+-activated ATPase and alkaline phosphatase of developing chick femora. Comp. Biochem. Physiol.51B, 19–24 (1975)

    Google Scholar 

  15. Post, R.L., Sen, A.K.: Sodium and potassium stimulated ATPase. Meth. Enzymol.10, 762–768 (1967)

    Google Scholar 

  16. Shami, Y., Radde, I.C.: Calcium-stimulated ATPase of guinea pig placenta. Biochim. Biophys. Acta249, 345–352 (1971)

    PubMed  Google Scholar 

  17. Taussky, H.H., Shorr, E.: A microcolorimetric method for the determination of inorganic phosphorus. J. Biol. Chem.22, 675–685 (1953)

    Google Scholar 

  18. Linhardt, K., Walter, K.: In Methods of Enzymatic Analysis (Bergmeyer, H.U. ed.), pp. 783–785. New York: Academic Press, 1965

    Google Scholar 

  19. Ghosh, N.K., Fishman, W.H.: Purification and properties of molecular weight variant of human placental alkaline phosphatase. Biochem. J.108, 779–792 (1968)

    PubMed  Google Scholar 

  20. Gothlin, G., Ericsson, J.L.E.: Fine structural localization of alkaline phosphomonoesterase in the fracture callus of the rat. Isr. J. Med. Sci.7, 488–490 (1971)

    Google Scholar 

  21. Razin, S.: Reconstitution of biological membranes, Biochim. Biophys. Acta265, 241–296 (1972)

    PubMed  Google Scholar 

  22. MacLennan, D.H.: Resolution of the calcium transport system of sarcoplasmic reticulum. Can. J. Biochem.53, 251–261 (1975)

    PubMed  Google Scholar 

  23. Fernley, H.N.: Mammalian alkaline phosphatase. In: The Enzymes (P.C. Boyer, ed.), Vol. 4, pp. 417–447. New York: Academic Press, 1971

    Google Scholar 

  24. Moss, D.W., Eaton, R.H., Smith, J.K., Whitby, L.G.: Association of inorganic-pyrophosphatase activity with human alkaline phosphatase preparations. Biochem. J.102, 53–57 (1967)

    PubMed  Google Scholar 

  25. Fernley, H.N., Walker, P.G.: Studies on alkaline phosphatase. biochem. J.104, 1011–1018 (1967)

    PubMed  Google Scholar 

  26. Linde, A., Granstrom, G., Magnusson, B.L.: Ca2+-ATPase in hard tissue forming cells. Calcif. Tissue Res. Suppl.21, 108–112 (1976)

    Google Scholar 

  27. Emmelot, P., Bos, C.J.: Studies of plasma membrane. V. On the lipid dependency of some phosphohydrolases of isolated rat liver plasma membrane. Biochim. Biophys. Acta150, 341–353 (1968)

    PubMed  Google Scholar 

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Author notes

  1. M. K. Guo

    Present address: School of Dentistry, College of Medicine, National Taiwan University, 100, Taipei, Taiwan

Authors and Affiliations

  1. Division of Oral Biology, School of Dentistry, University of Minnesota, 55455, Minneapolis, Minnesota, USA

    M. K. Guo & H. H. Messer

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  1. M. K. Guo
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  2. H. H. Messer
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Guo, M.K., Messer, H.H. A comparison of Ca2+-, Mg2+-ATPase and alkaline phosphatase activities of rat incisor pulp. Calc. Tis Res. 26, 33–38 (1978). https://doi.org/10.1007/BF02013231

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  • DOI: https://doi.org/10.1007/BF02013231

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