1887

Abstract

The addition of nitrite, the product of the reaction catalysed by nitrate reductase, to cell suspensions of the yeast caused a reversible inactivation of NADPH-dependent nitrate reductase activity. The haem- and Mo-dependent and Mo-dependent activities of nitrate reductase, determined with the non-physiological electron donors FMNHand reduced methyl viologen respectively, were less affected. A similar inactivation was found with the proton ionophores 2,4-dinitrophenol and carbonyl cyanide -chlorophenylhydrazone. The inactive enzyme was found in the particulate fraction and cosedimented with the mitochondrial fraction. When the NADPH-dependent nitrate reductase activity was restored the enzyme was found in the soluble fraction. The inactivation of nitrate reductase by nitrite, 2,4-dinitrophenol and carbonyl cyanide -chlorophenylhydrazone was dependent on the external pH. The treatment of isolated mitochondria at alkaline pH with Triton X-100 solubilized about 30% of the inactive enzyme.

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1994-10-01
2021-10-19
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References

  1. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254
    [Google Scholar]
  2. Dubois M.F., Hovanessian A.G., Bensaude O. Heat- shock-induced denaturation of proteins. J Biol Chem 1991; 266:9707–9711
    [Google Scholar]
  3. Gething M.-J., Sambrook I. Protein folding in the cell. Nature 1992; 355:33–45
    [Google Scholar]
  4. González C., Siverio J.M. Effect of nitrogen source on the levels of nitrate reductase in the yeast Hansenula anómala. J Gen Microbiol 1992; 138:1445–1451
    [Google Scholar]
  5. Hipkin C.R. Nitrate assimilation in yeast. In Molecular and Genetic Aspects of Nitrate Assimilation 1989 Edited by Wray J.L., Kinghorn J.R. Oxford: Oxford Science Publications; pp 51–68
    [Google Scholar]
  6. Horau J., Nato A., Lavergne D., Flipo V., Hirel B. Nitrate reductase activity changes during a culture cycle of tobacco cells: the participation of a membrane-bound form enzyme. Plant Sci 1991; 79:193–204
    [Google Scholar]
  7. Mager W.H., Moradas-Ferreira P. Stress response of yeast. Biochem J 1993; 290:1–13
    [Google Scholar]
  8. Marzluf G.A. Regulation of sulfur and nitrogen metabolism in filamentous fungi. Annu Rep Microbiol 1993; 47:31–55
    [Google Scholar]
  9. Mazón M.J., Gancedo J.M., Gancedo C. Phosporyl- ation and inactivation of yeast fructose-bisphosphatase in vivo by glucose and by proton ionophores. Eur J Biochem 1982; 127:605–608
    [Google Scholar]
  10. Minagawa N., Yoshimoto A. Assimilatory nitrate reductase of Hansenula anómala: its electron donors and cellular distribution. Agric Biol Chem 1983; 47:125–127
    [Google Scholar]
  11. Minagawa N., Yoshimoto A. The in vivo inactive nitrate reductase from Hansenula anómala. Agrie Biol Chem 1984; 48:1907–1909
    [Google Scholar]
  12. Navarrete R., Serrano R. Solubilization of yeast plasma membranes and by different types of non-denaturing detergents. Biochim Biophys Acta 1983; 728:403–408
    [Google Scholar]
  13. Pichitony F., Méténier G. Régulation de la biosynthèse et localisation de la nitrate réductase d–Hansenula anomala. Ann Inst Pasteur 1967; 112:701–711
    [Google Scholar]
  14. Rickwood D., Wilson M.T., Darley-Usmar V.M. Isolation and characteristics of intact mitochondria. In Mitochondria 1987 Edited by Darley-Usmar V.M., Rickwood D., Wilson M.T. Oxford: IRL Press; a Practical Approach, pp 1–16
    [Google Scholar]
  15. Roberts C.J., Raymond C.K., Yamashiro C.T., Stevens T.H. Methods for studying the yeast vacuole. Methods Enzymol 1991; 194:644–661
    [Google Scholar]
  16. Silver W.S. Pyridine nucleotide-nitrate reductase from Hansenula anómala, a nitrate reducing yeast. J Bacterio1 1956; 73:241–246
    [Google Scholar]
  17. Siverio J.M., González C., Mendoza-Riquel A., Pérez M.-D., González G. Reversible inactivation and binding to mitochondria of nitrate reductase by heat shock in the yeast Hansenula anómala. FEBS Eett 1993; 318:153–156
    [Google Scholar]
  18. Solomonson L.P., Barber M.J. Assimilatory nitrate reductase: functional properties and regulation. Annu Rev Plant physiol Plant Mol Boil 1990; 41:225–253
    [Google Scholar]
  19. Stadtman E.R. Discovery of glutamine synthetase cascade. Methods Enzymol 1990; 182:793–809
    [Google Scholar]
  20. Zauner E., Dellweg H. Purification and properties of the assimilatory nitrate reductase from the yeast Hansenula anomala. Eur J Appl Microbiol Biotechnol 1983; 17:90–95
    [Google Scholar]
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