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Possible mechanism of inhibition of cartilage alkaline phosphatase by insulin

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Summary

The inhibition of alkaline phosphatase by insulin is a finding reported by many researchers but the mechanism of this inhibition has not been studied. Since alkaline phosphatase is an important factor in the mechanism of calcification and an impairment of mineralisation has been observed in diabetes mellitus, a study was carried out to assess the effect of the hormone on alkaline phosphatase measured in chondrocytes, in matrix vesicles and in a purified enzyme preparation. Enzyme activity was inhibited by insulin. The lowest active concentration was 10−6 M and maximal inhibition was obtained at about 10−4 M. The inhibition is of the uncompetitive type. Full recovery of the hormone-inhibited enzyme was obtained with 10−4 M 2-mercaptoethanol. Data suggest direct interaction between the alkaline phosphatase and insulin molecules, involving either disulfide cross linkages or the metal chelating activity of insulin.

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References

  1. Ali S. Y., Saidera S. W., Anderson H. C.: Isolation and characterization of calcifying matrix vesicles from epiphyseal cartilage — Proc. nat. Acad. Sci. (Wash.)67, 1513, 1970.

    Article  CAS  Google Scholar 

  2. Baici A.: The specific velocity plot — Europ. J. Biochem.119, 9, 1981.

    Article  PubMed  CAS  Google Scholar 

  3. Bonucci E.: Matrix vesicles: their role in calcification. In:Linde A. (Ed.): Dentin and Dentinogenesis. CRC Press Inc., Boca Raton/FL, 1984, vol. 1, p. 135.

    Google Scholar 

  4. Bradford M. M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding — Anal. Biochem.72, 248, 1976.

    Article  PubMed  CAS  Google Scholar 

  5. Catalan R. E., Martinez A. M., Aragones M. D., Miguel B. G., Robles A.: Insulin action on brain microvessels: effects on alkaline phosphatase — Biochem. biophys. Res. Commun.150, 583, 1988.

    Article  PubMed  CAS  Google Scholar 

  6. Corvol M. T., Dumontier M. F., Rappaport R., Guyda H., Posner B. I.: The effect of a slightly acidic somatomedin peptide (ILAs) on the sulphation of proteoglycans from articular and growth plate chondrocytes in culture — Acta endocrinol. (Kbh.)89, 263, 1978.

    CAS  Google Scholar 

  7. De Bernard B., Bianco P., Bonucci E., Costantini M., Lunazzi G. C., Martinuzzi P., Modricky C., Moro L., Panfili E., Pollesello P., Stagni N., Vittur F.: Biochemical and immunohistochemical evidence that in cartilage and alkaline phosphatase is a Ca2+-binding glycoprotein — J. Cell Biol.103, 1615, 1986.

    Article  PubMed  Google Scholar 

  8. Dixon M.: The graphical determination of Km and Ki — Biochem. J.129, 197, 1972.

    PubMed  CAS  Google Scholar 

  9. Eisenthal R., Cornish-Bowden: A new graphical procedure for estimating enzyme kinetic parameters — Biochem. J.139, 715, 1974.

    PubMed  CAS  Google Scholar 

  10. Fernley H. N.: Mammalian alkaline phosphatases. In:Boyer P. D. (Ed.): The Enzymes. Academic Press, New York-London, 1971; vol. 4, p. 417.

    Google Scholar 

  11. Hui S. L., Epstein S., Johnston C. C. Jr. A prospective study of bone mass in patients with type I diabetes — J. clin. Endocrinol.60, 74, 1985.

    CAS  Google Scholar 

  12. Levin M. E., Boisseau V. C., Avioli L. V.: Effects of diabetes mellitus on bone mass in juvenile and adult-onset diabetes — New Engl. J. Med.294, 241, 1976.

    Article  PubMed  CAS  Google Scholar 

  13. Levy J. R., Murray E., Manolagas S., Olefsky J. M.: Demonstration of insulin receptors and modulation of alkaline phosphatase activity by insulin in rat osteoblastic cells — Endocrinology119, 1786, 1986.

    PubMed  CAS  Google Scholar 

  14. Lineweaver H., Burk D.: The determination of enzyme dissociation constants — J. Amer. chem. Soc.56, 658, 1934.

    Article  CAS  Google Scholar 

  15. McNair P., Madsbad S., Christiansen C., Christensen M. S., Faber O. K., Binder C., Transbøl I.: Development of bone mineral loss in insulin-treated diabetics: a one-and-a-half year follow-up study in sixty patients — Europ. J. clin. Invest.11, 55, 1981.

    Article  PubMed  CAS  Google Scholar 

  16. Miki A., Tanaka Y., Ogata S., Ikehara Y.: Selective preparation and characterisation of membranous and soluble forms of alkaline phosphatase from rat tissues — Europ. J. Biochem.160, 41, 1986.

    Article  PubMed  CAS  Google Scholar 

  17. Santiago J. V., McAlister W. H., Rabzan S. K., Bussman Y., Haymond M. V., Shackelford G., Weldon V. V.: Decreased cortical thickness and osteopenia in children with diabetes mellitus — J. clin. Endocrinol. Metabol.45, 845, 1977.

    Article  CAS  Google Scholar 

  18. Shore R. M., Chesney R., Mazzes R. B., Rose P. G., Bargman G. J.: Osteopenia in juvenile diabetes — Calcif. Tissue Int.33, 455, 1981.

    Article  PubMed  CAS  Google Scholar 

  19. Weiss J. M., Henthorn P. S., Lafferty M. A., Slaughter C., Raducha M., Harris H.: Isolation and characterisation of a cDNA encoding a human liver/bone/kidney-type alkaline phosphatase — Biochemistry (Wash.)83, 7182, 1986.

    CAS  Google Scholar 

  20. Wiske P. S., Wentworth S. M., Norton J. A. Jr., Epstein S., Johnston C. C. Jr.: Evaluation of bone mass and growth in young diabetics — Metabolism31, 848, 1982.

    Article  PubMed  CAS  Google Scholar 

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Gazzarrini, C., Stagni, N., Pollesello, P. et al. Possible mechanism of inhibition of cartilage alkaline phosphatase by insulin. Acta diabet. lat 26, 321–327 (1989). https://doi.org/10.1007/BF02624644

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