Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1978 Apr;75(4):1979–1983. doi: 10.1073/pnas.75.4.1979

Biochemical mechanisms of glucose-6-phosphate dehydrogenase deficiency.

A Morelli, U Benatti, G F Gaetani, A De Flora
PMCID: PMC392466  PMID: 273924

Abstract

A solid-phase radioimmunoassay for human glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ 1-oxidoreductase; EC 1.1.1.49) was developed that allowed the specific activity of this enzyme protein to be measured in lysates from whole erythrocyte populations, in lysates from erythrocytes of different ages, and in purified samples. The enzyme was highly purified from erythrocytes of single donors by a simple procedure of affinity chromatography with insolubilized adenosine 2',5'-bisphosphate. These techniques were used in an attempt to elucidate the molecular mechanisms leading to deficiency of glucose-6-phosphate dehydrogenase activity in two genetic variants of the enzyme, i.e., the Mediterranean and the Seattle-like variants. The results indicate that the lowered activity of erythrocytes containing the Mediterranean variant of glucose-6-phosphate dehydrogenase is related to an enhanced rate of degradation of a catalytically defective protein synthesized at a nearly normal rate. Synthesis of a normally functioning protein and an increased breakdown of it are involved in the Seattle-like variant of the enzyme.

Full text

PDF
1979

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Babalola O., Cancedda R., Luzzatto L. Genetic variants of glucose 6-phosphate dehydrogenase from human erythrocytes: unique properties of the A - variant isolated from "deficient" cells. Proc Natl Acad Sci U S A. 1972 Apr;69(4):946–950. doi: 10.1073/pnas.69.4.946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CUNNINGHAM L. W., NUENKE B. J., STRAYHORN W. D. Sulfhydryl content and tryptic susceptibility of thermally denatured ovalbumin. J Biol Chem. 1957 Oct;228(2):835–845. [PubMed] [Google Scholar]
  3. Cohen P., Rosemeyer M. A. Human glucose-6-phosphate dehydrogenase: purification of the erythrocyte enzyme and the influence of ions on its activity. Eur J Biochem. 1969 Mar;8(1):1–7. doi: 10.1111/j.1432-1033.1969.tb00487.x. [DOI] [PubMed] [Google Scholar]
  4. Corash L. M., Piomelli S., Chen H. C., Seaman C., Gross E. Separation of erythrocytes according to age on a simplified density gradient. J Lab Clin Med. 1974 Jul;84(1):147–151. [PubMed] [Google Scholar]
  5. De Flora A., Lorenzoni I., Mangiarotti M. A., Dina D., Bonsignore A. Electrophoretic behaviour of human erythrocyte glucose 6-phosphate dehydrogenase during purification. Biochem Biophys Res Commun. 1968 May 10;31(3):501–507. doi: 10.1016/0006-291x(68)90505-6. [DOI] [PubMed] [Google Scholar]
  6. DeFlora A., Morelli A., Benatti U., Giuliano F. An improved procedure for rapid isolation of glucose 6-phosphate dehydrogenase from human erythrocytes. Arch Biochem Biophys. 1975 Jul;169(1):362–363. doi: 10.1016/0003-9861(75)90353-7. [DOI] [PubMed] [Google Scholar]
  7. GLOCK G. E., McLEAN P. Further studies on the properties and assay of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase of rat liver. Biochem J. 1953 Oct;55(3):400–408. doi: 10.1042/bj0550400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. KIRKMAN H. N., CROWELL B. B. Molecular deficiency of glucose-6-phosphate dehydrogenase in primaquine sensitivity. Nature. 1963 Jan 19;197:286–287. doi: 10.1038/197286a0. [DOI] [PubMed] [Google Scholar]
  9. Kahn A., Cottreau D., Boivin P. Molecular mechanism of glucose-6-phosphate dehydrogenase deficiency. Humangenetik. 1974;25(2):101–109. doi: 10.1007/BF00283310. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Lenzerini L., Meera Khan P., Filippi G., Rattazzi M. C., Ray A. K., Siniscalco M. Characterization of glucose-6-phosphate dehydrogenase variants. I. Occurrence of a G6PD Seattle-like variant in Sardinia and its interaction with the G6PD Mediterranean variant. Am J Hum Genet. 1969 Mar;21(2):142–153. [PMC free article] [PubMed] [Google Scholar]
  12. Luzzatto L. Genetic heterogeneity and pathophysiology of G6PD deficiency. Br J Haematol. 1974 Oct;28(2):151–155. doi: 10.1111/j.1365-2141.1974.tb06649.x. [DOI] [PubMed] [Google Scholar]
  13. MARKS P. A., JOHNSON A. B. Relationship between the age of human erythrocytes and their osmotic resistance: a basis for separating young and old erythrocytes. J Clin Invest. 1958 Nov;37(11):1542–1548. doi: 10.1172/JCI103746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morelli A., Benatti U. Simple chemical synthesis of a specific effector for the affinity chromatography of nicotinamide adenine dinucleotide phosphate-dependent dehydrogenases. Ital J Biochem. 1974 Sep-Oct;23(5):279–291. [PubMed] [Google Scholar]
  15. Morelli A., De Flora A. Isolation and partial charaterization of an NADP- and NADPH- binding protein from human erythrocytes. Arch Biochem Biophys. 1977 Mar;179(2):698–705. doi: 10.1016/0003-9861(77)90159-x. [DOI] [PubMed] [Google Scholar]
  16. Piomelli S., Corash L. M., Davenport D. D., Miraglia J., Amorosi E. L. In vivo lability of glucose-6-phosphate dehydrogenase in GdA- and GdMediterranean deficiency. J Clin Invest. 1968 Apr;47(4):940–948. doi: 10.1172/JCI105786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Piomelli S., Lurinsky G., Wasserman L. R. The mechanism of red cell aging. I. Relationship between cell age and specific gravity evaluated by ultracentrifugation in a discontinuous density gradient. J Lab Clin Med. 1967 Apr;69(4):659–674. [PubMed] [Google Scholar]
  18. Rosa R., Alexandre Y., Kaplan J. C., Dreyfus J. C. Comportement immunologique de la glucose-6-phosphate-deshydrogenase érythrocytaire chez des mutants déficients. Clin Chim Acta. 1970 Aug;29(2):209–214. doi: 10.1016/0009-8981(70)90038-0. [DOI] [PubMed] [Google Scholar]
  19. Yoshida A. Glucose 6-phosphate dehydrogenase of human erythrocytes. I. Purification and characterization of normal (B+) enzyme. J Biol Chem. 1966 Nov 10;241(21):4966–4976. [PubMed] [Google Scholar]
  20. Yoshida A., Stamatoyannopoulos G., Motulsky A. G. Negro variant of glucose-6-phosphate dehydrogenase deficiency (A-) in man. Science. 1967 Jan 6;155(3758):97–99. doi: 10.1126/science.155.3758.97. [DOI] [PubMed] [Google Scholar]
  21. de Flora A., Morelli A., Frascio M., Corte G., Curti B., Galliano M., Gozzer C., Minchiotti L., Mareni C., Gaetani G. Radioimmunoassay and chemical properties of glucose 6-phosphate dehydrogenase and of a specific NADP(H)-binding protein (FX) from human erythrocytes. Biochim Biophys Acta. 1977 Nov 7;500(1):109–123. doi: 10.1016/0304-4165(77)90051-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES