Metal Displacement Effects on Monoesterase Activity of Calf Intestinal Alkaline Phosphatase
Abstract
The mechanism of modulation of alkaline phosphatase activity by metal ions has not been fully elucidated. We investigated the time-dependent modulatory effects of Mg2+ and Zn2+ in promoting the hydrolysis of para-nitrophenyl phosphate (monoesterase reaction) by calf intestinal alkaline phosphatase (CIAP) and the effects of addition of the activating metal ions to the metal-inhibited enzyme. The CIAP was affected by changes in pre-incubation time in the presence of the two metal cofactors. Both Mg2+ (0.1 – 0.25 mM) and Zn2+ (0.1 – 5 mM) modulated Zn2+- and Mg2+- inhibited monoesterase activity of CIAP. The CIAP activity was inhibited when the enzyme was pre-incubated with 1 mM Ca2+. Further addition of Mg2+ (0.1 – 2 mM) did not completely restore the activity though partly relieved the inhibition caused by Ca2+- pre-incubated enzyme. Again, addition of 2mM Zn2+ to Ca2+- pre-incubated alkaline phosphatase completely restored the activity of the enzyme. This study suggests that Mg2+ and Zn2+ regulate the catalytic property of each other and modulate the inhibitory effect of Ca2+ in alkaline phosphatase catalysis through a displacement effect. The modulation of Ca2+-inhibited CIAP activity by Mg2+ and Zn2+ may be explored in the treatment of disorders of bone mineralization especially those arising from inhibited alkaline phosphatase activity.
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References
Ahlers, J. (1975). The mechanism of hydrolysis of beta glycerophosphate by kidney alkaline phosphatase. Biochemical Journal 149, 535-546.
Arise, R. O., Bolaji, F. F., Jimoh, O. A., Adebayo, J. O., Olorunniji, F. J. and Malomo, S. O. (2005). Regulatory effect of divalent cations on rat liver alkaline phosphatase activity: How Mg2+ activates (and inhibits) the hydrolysis of p-nitrophenylphosphate. Biokemistri 17: 129-136.
Bortolato, M., Besson, F. and Roux, B. (1999). Role of metal ions on the secondary and quaternary structure of alkaline phosphatase from bovine intestinal mucosa. Proteins: Structure, Function and Genetics 37: 310 - 318.
Bosron W. F., Anderson, R. A., Falk, M. C., Kennedy, F. S. and Valle, B. L. (1977). Effect of magnesium on properties of zinc alkaline phosphatase. Biochemistry 16: 610-614.
Ciancaglini, P., Pizauro, J. M., Grechi, M. J., Curti, C. and Leone, F. A. (1989). Effects of Zn2+ on phosphor hydrolytic activity of rat matrix – induced alkaline phosphatase. Cellular and Molecular Biology 35: 503-510.
Deen, R. L. (2002). Kinetic studies with alkaline phosphatase in the presence and absence of inhibitors and divalent cations. Biochemistry and Molecular Biology Education 30: 401 - 407.
Genge, B. R., Sauer, G. R., Wu, L. N. Y., Mclean, F. M. and Wuthier, R. E. (1988). Correlation between loss of alkaline phosphatase activity and accumulation of calcium during matrix vesicles-mediated mineralization. Journal of Biological Chemistry 263: 18513-18519.
Hessle, L., Johnson, K. A., Anderson, H. C., Narisawa, S., Sali, A., Goding, J. W., Terkerltaub, R. and Millan, J. L. (2002). Tissue-specific alkaline phosphatase and plasma cell membrane Glycoprotein-1 are central antagonistic regulators of bone mineralization. Proceedings of the National Academy of Science 99 (14): 9445 - 9449.
Holtz, K.M., Stec, B., and Kantrowitz, E.R. (1999). A model of the transition state in the alkaline phosphatase reaction. Journal of Biological Chemistry 274: 8351-8354.
Hoylaerts, M. F., Manes, T. and Millan, J. L. (1997). Mammalian alkaline phosphatases are allosteric enzymes. Journal of Biological Chemistry 272: 22781 - 22787.
Hung, H. and Chang, G. (2001). Differentiation of the slow binding mechanism for magnesium ion activation and zinc ion inhibition of human alkaline phosphatase. Protein science, 10: 34 – 45.
Igunnu, A., Osalaye, D. S., Olorunsogo, O. O., Malomo, S. O. and Olorunniji, F. J. (2011). Distinct metal ion requirements for the phosphomonoesterase and phosphodiesterase activities of calf intestinal alkaline phosphatase. The Open Biochemistry Journal 5: 45-50.
Kim, E. E. and Wyckoff, H. W. (1991). Reaction mechanism of alkaline phospatases based on crystal structures: Two-metal ion catalysis. Journal of Molecular Biology 218: 449-464.
Le Du, M. H., Stigbrand, T., Taussig, M. J., Menez, A. and Stura, E. A. (2001). Crystal structure of alkaline phosphatase from human placenta at 1.8 A resolution. Implication for a substrate specificity. Journal of Biological Chemistry 276: 9158-9165.
Millan, J. L. (2006). Alkaline phosphatases: Structure, substrate specificity and functional relatedness to other members of a large superfamily of enzymes. Purinergic Signalling 2: 335-341.
Moss, D. W. (1982). Alkaline phosphatase Isoenzymes. Clinical Chemistry 28 (10): 2007-2016.
Muginova, S. V., Zhavoronkova, A. M., Polyakov, A. E., and Shekhovtsova, T. V. (2007). Applications of alkaline phosphatases from different sources in pharmaceutical and chemical analysis for the determination of their cofactors; zinc and magnesium ions. Analytical Science 23: 357-363.
Murphy, J. E., Tibbitts, T. T. and Kantrowitz, E. R. (1995). Mutations at positions 153 and 328 in Escherichia coli alkaline phosphatase provide insight towards the structure and function of mammalian and yeast alkaline phosphatases. Journal of Biological Chemistry 253: 604-617.
Olorunniji, F. J., Igunnu, A., Adebayo, J. O., Arise, R. O. and Malomo, S. O. (2007). Cofactor interaction in the activation of tissue non-specific Alkaline phosphatase: Synergistic effects of Zn2+ and Mg2+ ions. Biokemistri 19: 43 – 48.
Orimo, H. (2010). The mechanism of mineralization and the role of alkaline phosphatase in health and disease. Journal of Nippon Medical School 77: 4 – 12.
Petitclerc, C. and Fecteau, C. (1977). Mechanism of action of Zn2+ and Mg2+ on rat placental alkaline phosphatases II. Studies on membrane bound phosphatase in tissue sections and in whole placenta. Canadian Journal of Biochemistry and Cell Biology 55: 474-478.
Simopoulos, T. T. and Jencks, W. P. (1994). Alkaline phosphatase is an almost perfect enzyme. Biochemistry 33: 10375-10380.
Sowadski, J. M., Handschumacher, M. D., Murthy, H. M. K., Foster, B. A. and Wyckoff, H. W. (1985). Refined structure of alkaline phosphatase from Escherichia coli at 2.8 Å resolution. Journal of Biological Chemistry 186: 417-433.
Zalatan, J. G., Fenn, T. D. and Hersclag, D. (2008). Comparative enzymology in the alkaline phosphatase superfamily to determine the catalytic role of an active site metal ion. Journal of Molecular Biology 384: 1174-1189.
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