1887

Abstract

SUMMARY: Glutathione reductase was purified to homogeneity from the unicellular alga The enzyme was a monomer with a molecular mass of 54-56 kDa as judged by gel filtration and SDS-PAGE. The activity was maximal at pH 8.2 and 49 °C. The enzyme was specific for NADPH, but not for NADH. The reverse reaction with NAD(P) and GSH (glutathione, reduced form) was not observed in the pH range 4.8-8.2. values for NADPH and GSSG (glutathione, oxidized form) were 10.6 μ;m and 54.1 μ;M, respectively. Thiol inhibitors and metal ions such as Hg and Cu markedly inhibited the enzyme activity. Activity was lost when the apoenzyme was prepared by dialysis, but was restored to 40% of the original activity by the addition of 50 μ;M-FAD. The enzyme reaction proceeded via a branching mechanism. Upon immunoprecipitation, glutathione reductase activity of was inhibited 50% and 90% by antibodies generated against spinach and glutathione reductases, respectively. Both antibodies cross-reacted with glutathione reductase in an immunoblot analysis.

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/content/journal/micro/10.1099/00221287-139-9-2233
1993-09-01
2021-10-16
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References

  1. Alscher R.G. 1989; Biosynthesis and antioxidant function of glutathione in plants.. Physiologic Plantarum 77:457–464
    [Google Scholar]
  2. Arscott L.D., Drake D.M., Williams C.H. Jr 1989; Inactivation-reactivation of two-electron reduced Escherichia coli glutathione reductase involving a dimer-monomer equilibrium.. Biochemistry 28:3591–3598
    [Google Scholar]
  3. Bradford M.M. 1976; A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.. Analytical Biochemistry 717:1448–1454
    [Google Scholar]
  4. Carlberg I., Mannervik B. 1975; Purification and characterization of the flavoenzyme glutathione reductase from rat liver.. Journal of Biological Chemistry 250:5475–5480
    [Google Scholar]
  5. Carlberg I., Mannervik B. 1985; Glutathione reductase.. Methods in Enzymology 113:484–495
    [Google Scholar]
  6. Foyer C., Lclandais M., Galap C., Kunert K.J. 1991; Effects of elevated cytosolic glutathione reductase activity on the cellular glutathione pool and photosynthesis in leaves under normal and stress conditions.. Plant Physiology 97:863–872
    [Google Scholar]
  7. Halliwell B., Foyer C.H. 1978; Properties and physiological function of a glutathione reductase purified from spinach leaves by affinity chromatography.. Planta 139:9–17
    [Google Scholar]
  8. Halliwell , Gutteridge J.M.C. 1985 Free Radicals in Biology and Medicine. Oxford:: Oxford University Press.;
    [Google Scholar]
  9. Hurn B.A.L., Chantler S.M. 1980; Production of reagent antibodies.. Methods in Enzymology 70:104–142
    [Google Scholar]
  10. Kow Y.W., Smyth D.A., Gibbs M. 1982; Oxidation of reduced pyridine nucleotide by a system using ascorbate and hydrogen peroxide from plants and algae.. Plant Physiology 69:72–76
    [Google Scholar]
  11. Madamanchi N.R., Anderson J.V., Alscher R.G., Cramer C.L., Hess J.L. 1992; Purification of multiple forms of glutathione reductase from pea (Pisum sativumL.) seedlings and enzyme levels in ozone-fumigated pea leaves.. Plant Physiology 100:138–145
    [Google Scholar]
  12. Massey V., Williams C.H. 1965; On the reaction mechanism of yeast glutathione reductase.. Journal of Biological Chemistsy 240:4470–4480
    [Google Scholar]
  13. Meister A., Anderson M.E. 1983; Glutathione.. Annual Review of Biochemistry 52:711–760
    [Google Scholar]
  14. Shigeoka S., Nakano Y. 1991; Characterization and molecular properties of 2-oxoglutarate decarboxylase from Euglena gracilis.. Archives of Biochemistry and Biophysics 288:22–28
    [Google Scholar]
  15. Shigeoka S., Nakano Y., Kitaoka S. 1980; Metabolism of hydrogen peroxide in Euglena gracilis Z by L-ascorbic acid peroxidase.. Biochemical Journal 186:377–380
    [Google Scholar]
  16. Shigeoka S., Yasumoto R., Onishi T., Nakano Y., Kitaoka S. 1987a; Properties of monodehydroascorbate reductase and dehydroascorbate reductase and their participation in the regeneration of ascorbate in Euglena gracilis.. Journal of General Microhiology 133:227–232
    [Google Scholar]
  17. Shigeoka S., Onishi T., Nakano Y., Kitaoka S. l987b; Characterization and physiological function of glutathione reductase in Euglena gracilis Z.. Biochemical Journal 242:511–515
    [Google Scholar]
  18. Shigeoka S., Takeda T., Hanaoka T., Yokota A., Kitaoka S., Iizuka Y. 1990; Properties of selenium-induced glutathione peroxidase in low-CO2-grown Chlamydomonas reinhardtii.. Current Research in Photosynthesis 4:615–618
    [Google Scholar]
  19. Shigeoka S., Takeda T., Hanaoka T. 1991; Characterization and immunological properties of selenium-containing glutathione peroxidase induced by selenite in Chlamydomonas reinhardtii.. Bio-chemical Journal 275:623–627
    [Google Scholar]
  20. Smith I.K., Vierheller T.L., Thorne C.A. 1989; Properties and functions of glutathione reductase in plants.. Physiologia Plantarum 77:449–456
    [Google Scholar]
  21. Takeda T., Shigeoka S., Hirayama O., Mitsunaga T. 1992; The presence of enzymes related to glutathione metabolism and oxygen metabolism in Chlamydomonas reinhardtii.. Bioscience, Bio-technology, and Biochemistry 56:1662–1663
    [Google Scholar]
  22. Tanaka K., Saji H., Kondo N. 1988; Immunological properties of spinach glutathione reductase and inductive biosynthesis of the enzyme with ozone.. Plant and Cell Physiology 29:637–642
    [Google Scholar]
  23. Tel-Or E., Huflejt M.E., Packer L. 1986; Hydroperoxide metabolism in cyanobacteria.. Archives of Biochemistry and Bio-physics 246:396–402
    [Google Scholar]
  24. Thieme R., Pai E.F., Schirmer R.H., Schultz G.E. 1981; Three dimensional structure of glutathione reductase at two A resolution.. Journal of Molecular Biology 152:763–782
    [Google Scholar]
  25. Williams C.H. Jr 1976; Glutathione reductase.. Enzymes 13:89–173
    [Google Scholar]
  26. Worthington D.J., Rosemeyer M.A. 1974; Human glutathione reductase: purification of the crystalline enzyme from erythrocytes.. European Journal of Biochemistry 48:167–177
    [Google Scholar]
  27. Worthington D.J., Rosemeyer M.A. 1976; Glutathione reductase from human erythrocytes. Catalytic properties and aggregation.. European Journal of Biochemistry 67:231–238
    [Google Scholar]
  28. Yokota A., Shigeoka S., Onishi T., Kitaoka S. 1988; Selenium as induced of glutathione peroxidase in low-CO2-grown Chlamydomonas reinhardtii.. Plant Physiology 86:649–651
    [Google Scholar]
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