Skip to main content
Download PDF
  • Published:

Procedure for Blood Glutathione Peroxidase Determination in Cattle and Swine

Analyseprocedure til bestemmelse af glutathion peroxidase i kvæg-og svineblod.

Acta Veterinaria Scandinavica volume 23, pages 515–527 (1982)Cite this article

Abstract

An improved testing system has been developed for direct measurement of glutathione peroxidase activity in heparinized whole blood at 37°C. Without loss in net yield the consumption of reagents has been found to be considerably lower with the new technique than with previously described techniques. Within the range 0–700 mKat/1 the GSH-Px activity in red cells may be measured with a high degree of accuracy and reproducibility without preceding separation and washing. The stability of the enzyme in bovine and porcine whole blood at 22°C, 4°C, and —20°C was determined.

Sammendrag

En analysemetode til bestemmelse af glutathion peroxidase i heparinstabiliseret fuldblod fra kvæg og svin er optimeret med hen-syn til pH, ionstyrke og donor- og acceptorsubstrat-koncentrationer ved 37°C. I forhold til tidligere beskrevet teknik er reagensforbruget reduceret uden fald i nettoudbyttet. I intervallet 0–700 mKat/1 måles GSH-Px aktiviteten i erytrocyter uden forudgående separations- og vaskeprocedurer. Enzymets stabilitet i fuldblod fra kvaeg og svin er fastlagt ved 22°C, 4°C og — 20°C.

References

  • Allen, W. M., W. H. Parr, P. H. Anderson, S. Berrett, R. Bradley & D. S. P. Patterson: Selenium and the activity of glutathione peroxidase in bovine erythrocytes. Vet. Rec. 1975, 102, 360–361.

    Article  Google Scholar 

  • Anderson, P. H., S. Berrett & D. S. P. Patterson: Glutathione peroxidase activity in erythrocytes and muscle of cattle and sheep and its relationship to selenium. J. comp. Path. 1978, 88, 181–189.

    Article  CAS  PubMed  Google Scholar 

  • Chavez, E. R.: Effect of dietary selenium on glutathione peroxidase activity in piglets. Ganad. J. Anim, Sci. 1979a, 59, 67–75.

    Article  CAS  Google Scholar 

  • Chavez, E. R.: Effect of dietary selenium depletion and repletion on plasma glutathione peroxidase activity and selenium concentration in blood and body tissue of growing pigs. Ganad. J. Anim. Sci. 1979b, 59, 761–771.

    Article  CAS  Google Scholar 

  • Combs, G. F., T. Noguchi & M. L. Scott: Mechanisms of action of selenium and vitamin E in protection of biological membranes. Fed. Proc. 1975, 34, 2090–2095.

    CAS  PubMed  Google Scholar 

  • Flohé, L. & I. Brand: Some hints to avoid pitfalls in quantitative determination of glutathione peroxidase (E.G. 1.11.1.9). Z. Klin. Chem. Klin. Biochem. 1970, 8, 156–161.

    PubMed  Google Scholar 

  • Flohé, L., W. Günzler, G. Jung, E. Schaich & F. Schneider: Glutathion-Peroxidase. II. Substratspezifitát und Hemmbarkeit durch Sub-stratanaloge. (Glutathione peroxidase. II. Substrate specificity and inhibition by substrate analogues). Hoppe-Seylers Z. Physiol. Chem. 1971, 352, 159–169.

    Google Scholar 

  • Flohé, L., W. A. Giinzler & H. H. Schock: Glutathione peroxidase. A selenoenzyme. FEBS Letters 1973, 32, 132–134.

    Article  PubMed  Google Scholar 

  • Giinzler, W. A., H. Kr enter s & L. Flohé: An improved coupled test procedure for glutathione peroxidase (E.G. 1.11.1.9) in blood. Z. Klin. Chem. Klin. Biochem. 1974, 12, 444–448.

    Google Scholar 

  • Giinzler, W. A., M. Vergin, I. Müller & L. Flohé: Glutathion-Peroxidase. VI. Die Reaktion der Glutathion-Peroxidase mit verschiedenen Hydroperoxiden. (Glutathione peroxidase. VI. The reaction of glutathione peroxidase with different hydroperoxides). Hoppe-Seylers Z. Physiol. Chem. 1972, 353, 1001–1004.

    Google Scholar 

  • Hoekstra, W. G.: Biochemical function of selenium and its relation to vitamin E. Fed. Proc. 1975, 34, 2083–2089.

    CAS  PubMed  Google Scholar 

  • Hussein, K. S. M. & B. E. V. Jones: Effects of different anticoagulants on the determination of erythrocyte glutathione peroxidase. Acta vet. scand. 1981, 22, 472–479.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kraus, R. J. & H. E. Ganther: Reaction of cyanide with glutathione peroxidase. Biochem. Biophys. Res. Commun. 1980, 96, 1116–1122.

    Article  CAS  PubMed  Google Scholar 

  • Lineweaver, H. & D. Burk: The determination of enzyme dissociation constant. J. Amer. chem. Soc. 1934, 56, 658–666.

    Article  CAS  Google Scholar 

  • Little, C.: Steroid hydroperoxides as substrates for glutathione peroxidase. Biochem. Biophys. Acta 1972, 284, 375–381.

    CAS  PubMed  Google Scholar 

  • Little, C. & P. J. O’Brien: An intracellular GSH-peroxidase with a lipid peroxidase substrate. Biochem. Biophys. Res. Commun. 1968, 31, 145–150.

    Article  CAS  PubMed  Google Scholar 

  • Little, C., O. Olinescu, K. G. Reid & P. J. O’Brien: Properties and regulation of glutathione peroxidase. J. Biol. Chem. 1970, 245, 3632–3636.

    CAS  PubMed  Google Scholar 

  • Moss, D. W.: Nomenclature and units in enzymology. In: Principles of Enzymatic Analysis. Ed. H. U. Bergmeyer. Verlag Chemi 1978.

  • Noguchi, T., A. H. Cantor & M. L. Scott: Mode of action of selenium and vitamin E in prevention of exudative diathesis in chicks. J. Nutr. 1973, 103, 1502–1511.

    Article  CAS  PubMed  Google Scholar 

  • Oh, S. H., H. E. Ganther & W. G. Hoekstra: Selenium as a component of glutathione peroxidase isolated from ovine erythrocytes. Biochemistry 1974, 13, 1825–1829.

    Article  CAS  PubMed  Google Scholar 

  • Omaye, S. T. & A. Tappel: Effect of dietary selenium on glutathione peroxidase in the chick. J. Nutr. 1974, 10b, 747–753.

  • Prohaska, J. R., S. H. Oh, W. G. Hoekstra & H. E. Ganther: Glutathione peroxidase: Inhibition by cyanide and release of selenium. Biochem. Biophys. Res. Commun. 1977, 74, 64–71.

    Article  CAS  PubMed  Google Scholar 

  • Rotruck, J. T., A. L. Pope, H. E. Ganther, A. B. Swanson, D. G. Hafeman & W. G. Hoekstra: Selenium: Biochemical role as a component of glutathione peroxidase. Science 1973, 179, 588–590.

    Article  CAS  PubMed  Google Scholar 

  • Scott, M. L., T. Noguchi & G. F. Combs: New evidence concerning mechanisms of action of vitamin E and selenium. Vitam. Horm. 1974, 32, 429–443.

    Article  CAS  PubMed  Google Scholar 

  • Spencer, N., D. A. Hopkinson & H. Harris: Adenosine deaminase polymorphism in man. Ann. Hum. Genet. 1968, 32, 9–14.

    Article  Google Scholar 

  • Stadtman, T. C: Selenium-Dependent enzymes. Ann. Rev. Biochem. 1980, 49, 93–110.

    Article  CAS  PubMed  Google Scholar 

  • Sunde, R. A. & W. G. Hoekstra: Structure, synthesis and function of glutathione peroxidase. Nutr. Rev. 1980, 38, 265–273.

    Article  CAS  PubMed  Google Scholar 

  • Underwood, E. J.: Trace Elements in Human and Animal Nutrition. 4. ed. Academic Press, New York 1977.

  • Van Vleet, J. F.: Current knowledge of selenium-vitamin E deficiency in domestic animals. J. Amer. vet. med. Ass. 1980, 176, 321–325.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. State Veterinary Serum Laboratory, Rolighedsvej 25, Copenhagen, DK-1958, Denmark

    N. Agergaard & P. Thode Jensen

Authors
  1. N. Agergaard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Thode Jensen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Agergaard, N., Jensen, P.T. Procedure for Blood Glutathione Peroxidase Determination in Cattle and Swine. Acta Vet Scand 23, 515–527 (1982). https://doi.org/10.1186/BF03546770

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1186/BF03546770

Keywords

Acta Veterinaria Scandinavica

ISSN: 1751-0147