Biochemical Genetics

, Volume 1, Issue 4, pp 359–371 | Cite as

Purification and characterization of genetic variants of 6-phosphogluconate dehydrogenase

  • Ling-yu Shih
  • Parvin Justice
  • David Yi-Yung Hsia


This paper describes the physicochemical characteristics in partially purified enzyme on subjects with the Pd A, Pd AB, and Pd B variants of 6-phosphogluconic dehydrogenase (6PGD). For these studies, whole blood was purified about 225-fold using ion exchange chromatography on DEAE cellulose column and fractionation with ammonium sulfate. 6PGD emerges as a single peak between 0.01 m and 0.1 m phosphate buffer on the column and is precipitated in the 55–80% fraction of ammonium sulfate. This purified enzyme can be stored frozen for several months without appreciable loss of activity and contains no detectable activity of glucose 6-phosphate dehydrogenase and glutathione reductase. The three variants of partially purified 6PGD varied from each other in two respects. The transitional temperature is 47.8 C for Pd A, 45.4 C for Pd AB, and 41.1 C for Pd B. The Km for 6PGA is 30 μm for Pd A, 21 μm for Pd AB, and 15 μmfor Pd B. These observations add further strength to the concept that the polymorphism in 6PGD represents alterations in either the configuration or structure of the protein molecule itself.


Cellulose Glutathione Glutathione Reductase Protein Molecule Single Peak 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bowman, J. E., Carson, P. E., Frischer, H., and deGaray, A. L. (1966). Genetics of starch-gel electrophoretic variants of human 6-phosphogluconic dehydrogenase: Population and family studies in the United States and in Mexico. Nature 210 811.Google Scholar
  2. Carson, P. E., Ajmar, F., Hashimoto, F., and Bowman, J. E. (1966). Electrophoretic demonstration of stromal effect on hemolysate glucose-6-phosphate dehydrogenase and 6-phosphogluconic dehydrogenase. Nature 210 813.Google Scholar
  3. Carter, N. D., Gould, S. R., Parr, C. W., and Walter, P. H. (1966). Differential inhibition of human red cell phosphogluconate dehydrogenase variants. Biochem. J. 97: 17P.Google Scholar
  4. Davidson, R. G. (1967). Electrophoretic variants of human 6-phosphogluconate dehydrogenase: Population and family studies and description of a new variant. Ann. Human Genet. 30 355.Google Scholar
  5. Fildes, R. A., and Parr, C. W. (1963). Human red-cell phosphogluconate dehydrogenase. Nature 200 890.Google Scholar
  6. 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. 55 400.Google Scholar
  7. Gordon, H., Keraan, M. M., and Vooijs, M. (1967). Variants of 6-phosphogluconate dehydrogenase within a community. Nature 214 466.Google Scholar
  8. Horecker, B. L., and Smyrniotis, P. Z. (1951). Phosphogluconic acid dehydrogenase from yeast. J. Biol. Chem. 193 371.Google Scholar
  9. Ingram, V. M. (1958). Abnormal human haemoglobins. I. Comparison of normal human and sicklecell haemoglobins by finger-printing. Biochim. Biophys. Acta 28 539.Google Scholar
  10. Ingram, V. M. (1963). The Hemoglobins in Genetics and Evolution. Columbia University Press, New York.Google Scholar
  11. Kirkman, H. N., and Hendrickson, E. M. (1963). Sex-linked electrophoretic difference in glucose-6-phosphate dehydrogenase. Am. J. Human Genet. 15 241.Google Scholar
  12. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin Phenol Reagent. J. Biol. Chem. 193 265.Google Scholar
  13. Luzzatto, L., and Allan, N. C. (1965). Different properties of glucose-6-phosphate dehydrogenase from human erythrocytes with normal and abnormal enzyme levels. Biochem. Biophys. Res. Comm. 21 547.Google Scholar
  14. Parr, C. W. (1966). Erythrocyte phosphogluconate dehydrogenase polymorphism. Nature 210 487.Google Scholar
  15. Parr, C. W., and Fitch, L. I. (1967). Inherited quantitative variations of human phosphogluconate dehydrogenase. Ann. Human Genet. 30 339.Google Scholar
  16. Parr, C. W., and Parr, I. B. (1965). Stability differences of inherited variants of human red cell phosphogluconate dehydrogenase. Biochem. J. 95: 16P.Google Scholar
  17. Tashian, R. E., Rigg, S. K., and Yu, Y.-S. (1966). Characterization of a mutant human erythrocyte carbonic anhydrase: carbonic anhydrase IcGuam. Arch. Biochem. Biophys. 117 320.Google Scholar
  18. Yoshida, A. (1967). A single amino acid substitution (asparagine to aspartic acid) between normal (B+) and the common Negro variant (A+) of human G-6-P D. Proc. Natl. Acad. Sci. 67 835.Google Scholar
  19. Zinkham, W. J., and Lenhard, R. E. J. (1959). Metabolic abnormalities of erythrocytes from patients with congenital nonspherocytic hemolytic anemia. J. Ped. 55 319.Google Scholar

Copyright information

© Plenum Publishing Corporation 1968

Authors and Affiliations

  • Ling-yu Shih
    • 1
  • Parvin Justice
    • 1
  • David Yi-Yung Hsia
    • 1
  1. 1.Genetic Clinic of the Children's Memorial Hospital and the Department of PediatricsNorthwestern University Medical SchoolChicago

Personalised recommendations