Abstract
6-Phosphogluconate dehyrogenase is evident at all developmental stages of Drosophila melanogaster. The activity level is highest in early third instar larvae and declines to a lower, but relatively constant, level at all later stages of development. The enzyme is localized in the cytosolic portion of the cell. The A-isozymic form of 6-phosphogluconate dehydrogenase was purified to homogeneity and has a molecular weight of 105,000. The enzyme is a dimer consisting of subunits with molecular weights of 55,000 and 53,000. For the oxidative decarboxylation of 6-phosphogluconate the Km for substrate is 81 µm while that for NADP+ is 22.3 µm. The optimum pH for activity is 7.8 while the optimum temperature is 37 C.
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Betts, S. A., and Mayer, R. J. (1975) Purification and properties of 6-phosphogluconate dehydrogenase from rabbit mammary gland. Biochem. J. 151263.
Bewley, G. C., and Lucchesi, J. C. (1975). Lethal effects of low and “null” activity alleles of 6-phosphogluconate hydrogenase in Drosophila melanogaster. Genetics 79451.
Bowman, J. T., and Simmons, J. R. (1973). Gene modulation in Drosophila: Dosage compensation of Pgd + and Zw + genes. Biochem. Genet. 10319.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72248.
Browder, L. W., and Williamson, J. H. (1976) The effects of cinnamon on xanthine dehydrogenase, aldehyde oxidase and pyridoxal oxidase activity during development in Drosophila melanogaster. Dev. Biol. 53241.
Dyson, J. E. D., and D'Orazio, R. E. (1973). Sheep liver 6-phosphogluconate dehydrogenase: Inhibition by nucleotide phosphates and by other metabolic intermediates. J. Biol. Chem. 2485428.
Dyson, J. E. D., D'Orazio, R. E., and Hanson, W. H. (1973). Sheep liver 6-phosphogluconate dehydrogenase: Isolation procedure and effect of pH, ionic strength and metal ions on the kinetic parameters. Arch. Biochem. Biophys. 154623.
Geer, B. W., Bowman, J. T., and Simmons, J. R. (1974). The pentose shunt in wild-type and glucose-6-phosphate dehydrogenase deficient Drosophila melanogaster. J. Exp. Zool. 18777.
Geer, B. W., Kamiak, S. N., Kidd, K. R., Nishimura, R. A., and Yemm, S. J. (1976). Regulation of the oxidative NADP-enzyme tissue levels in Drosophila melanogaster. I. Modulation by dietary carbohydrate and lipid. J. Exp. Zool. 19515.
Geer, B. W., Woodward, C. G., and Marshall, S. D. (1978). Regulation of the oxidative NADP-enzymes tissue levels in Drosophila melanogaster. II. The biochemical basis of dietary carbohydrate and d-glycerate modulation. J. Exp. Zool. 203391.
Geer, B. W., Krochko, D., and Williamson, J. H. (1979a). Ontogeny, cell distribution and the physiological role of NADP-malic enzyme in Drosophila melanogaster. Biochem. Genet. 17867.
Geer, B. W., Krochko, D., Oliver, M. J., Walker, V. K., and Williamson, J. H. (1979b). A comparative study of the NADP-malic enzymes from Drosophila and chick liver. J. Exp. Zool. (in press).
Gerasimova, T. I., Kogan, G. L., Rosovskii, Ya. M., and Gvozdev, V. A. (1978). Study of genetic structure of Pgd and Zw loci coding for 6-phosphogluconate dehydrogenase (6PGD) and glucose 6-phosphate dehydrogenase in Drosophila melanogaster, Proc. 14th Int. Congr. Genet., Moscow, USSR, p. 14.
Glock, G. E., and McLean, P. (1953). Further studies on the properties and assay of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase of rat liver. Biochem. J. 55400.
Gvozdev, V. A., Gerasimova, T. I., Kogan, G. L., and Braslavskaya, O. Yu. (1976). Role of the pentose phosphate pathway in metabolism of Drosophila melanogaster elucidated by mutations affecting glucose 6-phosphate and 6-phosphogluconate dehydrogenase. FEB Lett. 6485.
Gvozdev, V. A., Gerasimova, T. I., Kogan, G. L. and Rosovsky, J. M. (1977). Investigations on the organization of genetic loci in Drosophila melanogaster: Lethal mutations affecting 6-phosphogluconate dehydrogenase and their suppression. Mol. Gen. Genet. 153191.
Horie, Y. (1967). Dehydrogenase in carbohydrate metabolism in larvae of the silkworm, Bombyx mori, L. L. J. Insect. Physiol. 131163.
Hughes, M. B., and Lucchesi, J. C. (1977). Genetic rescue of a lethal “null” activity allele of 6-phosphogluconate dehydrogenase in Drosophila melanogaster. Science 1971114.
Kazazian, H. H., Jr. (1966). Molecular size studies on 6-phosphogluconate dehydrogenase. Nature 212197.
Kogen, G. V., Rosovsky, Y. M., and Gvozdev, V. A. (1977). Purification and immunological characteristics of wild type and mutant forms of 6-phosphogluconate dehydrogenase from Drosophila melanogaster (in Russian). Mol. Biol. 111388.
Lewis, E. G. (1960). A new standard food medium. Drosophila Inform. Serv. 34117.
Lucchesi, J. C. (1973). Dosage compensation in Drosophila. Ann. Rev. Genet. 7225.
Lucchesi, J. C. (1977). Dosage compensation: Transcription-level regulation of X-linked genes in Drosophila. Am. Zool. 17685.
Ochoa, S. (1955). Malic enzyme. In Colowick, S. P., and Kaplan, N. O. (eds.), Methods in Enzymology, Vol. 1, Academic Press, New York, pp. 739–753.
Pearse, B. M. F., and Rosemeyer, M. A. (1974). The molecular weight and subunit structure of human erythrocyte 6-phosphogluconate dehydrogenase. Eur. J. Biochem. 42225.
Procsal, D. and Holten, D. (1972). Purification and properties of rat liver 6-phosphogluconate dehydrogenase: Activity at normal in vivo concentration of coenzyme. Biochemistry 111310.
Rippa, M., and Signorini, M. (1975). 6-Phosphogluconate dehydrogenase from Candida utilis. In Wood, W. A. (ed.), Methods in Enzymology, Vol. 41B, Academic Press, New York, pp. 237–240.
Rippa, M., Signorini, M., and Pontremoli, S. (1967). Purification and properties of two forms of 6-phosphogluconate dehydrogenase from Candida utilis. Eur. J. Biochem. 1170.
Rippa, M., Signorini, M., Bellini, T., Dallocchio, F. (1978). The active site of 6-phosphogluconate dehydrogenase: A phosphate binding site and its surroundings. Arch. Biochem. Biophys. 189516.
Seecof, R. L., Kaplan, W. D. and Futch, D. G. (1969). Dosage compensation for enzyme activities in Drosophila melanogaster. Proc. Natl. Acad. Sci. 62528.
Simcox, P. D., and Dennis, D. T. (1978). 6-Phosphogluconate dehydrogenase isoenzymes from the developing endosperm of Ricinus communis L. Plant Physiol. 62287.
Veeger, C., DerVartanian, D. V., and Zeylemaker, W. P. (1969). Succinate dehydrogenase. In Lowenstein, J. M. (ed.), Methods in Enzymology, Vol. 8, Academic Press, New York, pp. 81–90.
Weber, K., Pringle, J. R., and Osborn, M. (1972). Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. In Hirs, C. H. W., and Timasheff, S. N. (eds., Methods of Enzymology, Vol. 26, Academic Press, New York, pp. 3–28.
Young, W. J. (1966). X-linked electrophoretic variation in 6-phosphogluconate dehydrogenase in Drosophila melanogaster. J. Hered. 5758.
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This work was supported by National Research Council of Canada Grant A5860 and by the University of Calgary Research Policy and Grants Committee.
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Williamson, J.H., Krochko, D. & Geer, B.W. 6-Phosphogluconate dehydrogenase from Drosophila melanogaster. I. Purification and properties of the a isozyme. Biochem Genet 18, 87–101 (1980). https://doi.org/10.1007/BF00504362
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DOI: https://doi.org/10.1007/BF00504362