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Free Radicals Promote “In Vitro” a Different Intracellular Decay of Rabbit Reticulocyte and Erythrocyte Glycolytic Enzymes

  • Vilberto Stocchi
  • Beatrice Biagiarelli
  • Linda Masat
  • Francesco Palma
  • Fulvio Palma
  • Giovanni Piccoli
  • Luigi Cucchiarini
  • Mauro Magnani
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 307)

Summary

Rabbit red blood cells (RBC) were exposed in vitro to an oxygenradical-generating system represented by iron and acorbic acid. Under these experimental conditions we have investigated the effect of this system on some intracellular rabbit reticulocyte and erythrocyte enzymes. The results obtained have shown a pronounced decay of hexokinase activity both in the erythrocytes and reticulocytes when exposed to these radical species. We have found that the amount of hexokinase inactivated is at least three times higher in a blood sample with a percentage of reticulocytes of 50–60%. This different behaviour of the hexokinase decay in the erythrocytes and reticulocytes could be due to its different intracellular distribution related to the two distinct cells. In addition we have evaluated some important intracellular compounds involved in maintaining the redox and the energetic state of the cell such as the reduced glutathione and the adenine nucleotides and their degradation products, in order to understand if there is any correlation between the hexokinase decay and a change concerning the metabolic conditions of the rabbit reticulocytes and erythrocytes exposed to free radicals.

Keywords

Glycolytic Enzyme Energetic State Triose Phosphate Isomerase Hexokinase Activity Glyceraldehyde Phosphate 
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.

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References

  1. 1.
    R. P. Hebbel, J. W. Eaton, J. W. Balasingan and M. H. Steinberg, Spontaneous oxygen radical generation by sickle erythrocytes, J. Clin. Invest. 70:1253 (1982).PubMedCrossRefGoogle Scholar
  2. 2.
    G. Cohen and P. Hochstein, Generation of hydrogen peroxide in erythrocytes by hemolytic agents, Biochemistry 3:895 (1964).PubMedCrossRefGoogle Scholar
  3. 3.
    P. Hochstein and S. K. Jain, Association of lipid peroxidation and polymerization of membrane proteins with erythrocyte aging, Fed. Proc. Fed. Am. Soc. Exp. Biol. 40:183 (1981).Google Scholar
  4. 4.
    K. J. A. Davies, Free radicals and protein degradation in human red blood cells, in: “Cellular and Metabolic Aspects of Aging: The Red Cell as a Model” J. W. Eaton, D. K. Konzen and J. G. White, Alan R. Liss, Inc. New York.Google Scholar
  5. 5.
    K. J. A. Davies, The role of intracellular proteolytic systems in antioxydant defenses, in: “Superoxide and Superoxide Dismutase in Chemistry, Biology and Medicine” G. Rotilio, ed) Elsevier North-Holland Biomedical Press, Amsterdam.Google Scholar
  6. 6.
    K. J. A. Davies and A. L. Goldberg, Oxygen radicals stimulate intracellular proteolysis and lipid peroxidation by independent mechanisms in erythrocytes, J. Biol. Chem. 262:8220 (1987).PubMedGoogle Scholar
  7. 7.
    K. J. A. Davies and A. L. Goldberg, Protein damaged by oxigen radicals are rapidly degraded in extracts of red blood cells, J. Biol. Chem. 262:8227 (1987).PubMedGoogle Scholar
  8. 8.
    K. J. A. Davies, Intracellular proteolytic systems may function as secondary antioxidant defenses: an hypothesis, J. Free Radical in Biol. Med. 2:155 (1987).CrossRefGoogle Scholar
  9. 9.
    K. J. A. Davies, Protein damage and degradation by oxigen radical. I General aspects, J. Biol. Chem. 262:9895 (1987).PubMedGoogle Scholar
  10. 10.
    K. J. A. Davies, M. E. Delsignore and S. W. Lin, Protein damage and degradation by oxigen radicals. II Modification of amino acids, J. Biol. Chem. 262:9902 (1987).PubMedGoogle Scholar
  11. 11.
    K. J. A. Davies and M. E. Delsignore, Protein damage and degradation by oxygen radicals. III Modification of secondary and tertiary structure, J. Biol. Chem. 262:9908 (1987).PubMedGoogle Scholar
  12. 12.
    K. J. A. Davies, S. W. Lin and R. E. Pacifici, Protein damage and degradation by oxigen radicals. IV Degradation of denatured protein, J. Biol. Chem. 262:9914 (1987).PubMedGoogle Scholar
  13. 13.
    V. Stocchi, M. Magnani, F. Canestrari, M. Dachà and G. Fornaini, Rabbit red blood cell hexokinase evidence for two distinct forms, and their purification and characterization from reticulocytes, J. Biol. Chem. 256:7856 (1981).PubMedGoogle Scholar
  14. 14.
    E. Beutler, Part III. Red cell glycolytic enzymes, in: Red Cell Metabolism. “A manual of biochemical methods”, Grune & Stratton, New York.Google Scholar
  15. 15.
    V. Stocchi, M. Magnani, G. Piccoli and G. Fornaini, Hexokinase microheterogeneicity in rabbit red blood cells and its behaviour during reticulocytes maturation, Mol, cell. Biochem. 79:133 (1988).CrossRefGoogle Scholar
  16. 16.
    D. C. Salo, R. E. Pacifici, S. W. Lin, C. Giulivi and K. J. A. Davies, Superoxide dismutase undergoes proteolysis and fragmentation following oxidative modification and inactivation, J. Biol. Chem. 265:11919 (1990).PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Vilberto Stocchi
    • 1
  • Beatrice Biagiarelli
    • 1
  • Linda Masat
    • 1
  • Francesco Palma
    • 1
  • Fulvio Palma
    • 1
  • Giovanni Piccoli
    • 1
  • Luigi Cucchiarini
    • 1
  • Mauro Magnani
    • 1
  1. 1.Istituto di Chimica Biologica “Giorgio Fornaini”Università degli Studi di UrbinoUrbinoItaly

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