Summary
Nitrite reductase from barley seedlings was purified 50–60 fold by ammonium sulphate precipitation and gel filtration. No differences were established in the characteristics of nitrite reductases isolated in this way from either leaf or root tissues. The root enzyme accepted electrons from reduced methyl viologen, ferredoxin, or an unidentified endogenous cofactor. Enzyme activity in both tissues was markedly increased by growth on nitrate. This activity was not associated with sulphite reductase activity. Microbial contamination could not account for the presence of nitrite reductase activity in roots. Nitrite reductase assayed in vitro with reduced methyl viologen as the electron donor was inhibited by 2,4-dinitrophenol but not by arsenate.
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Abbreviations
- DNP:
-
2,4-dinitrophenol
- DEAE:
-
diethyl amino ethyl
References
Ahmad, J., Morris, I.: The effects of 2,4-dinitrophenol and other uncoupling agents on the assimilation of nitrate and nitrite by Chlorella. Biochim. biophys. Acta (Amst.) 162, 32–38 (1968).
Beevers, L., Hageman, R. H.: Nitrate reduction in higher plants. Ann. Rev. Plant Physiol. 20, 495–522 (1969).
Bollard, E. G.: Nitrogenous compounds in plant xylem sap. Nature (Lond.) 178, 1189–1190 (1956).
Bourne, W. F., Miflin, B. J.: An ATP dependent reduction of nitrate to ammonia by a cell free particulate system from barley roots. Biochem. biophys. Res. Commun. 40, 1305–1310 (1970).
Butt, V. S., Beeyers, H.: The regulation of pathways of glucose catabolism in maize roots. Biochem. J. 80, 21–27 (1961).
Hageman, R. H., Cresswell, C. F., Hewitt, E. J.: Reduction of nitrate, nitrite and hydroxylamine to ammonia by enzymes from higher plants. Nature (Lond.) 193, 247–250 (1962).
Hattori, A., Myers, J.: Reduction of nitrate and nitrite by subcellular preparations of Anabaena cydlindrica. Plant Physiol. 41, 1031–1036 (1966).
Hill, R., Bendall, F.: Crystallization of a photosynthetic reductase from a green plant. Nature (Lond.) 187, 417 (1960).
Hucklesby, D. P., Dalling, M. J., Hageman, R. H.: Some properties of two forms of nitrite reductase from corn (Zea mays L.) Scutellum. Planta (Berl.) 104, 220–233 (1972).
Joy, K. W., Hageman, R. H.: The purification of nitrite reductase from higher plants and its dependence on ferredoxin. Biochem. J. 100, 263–273 (1966).
Kemp, J. D., Atkinson, D. E., Ehret, A., Lazzarini, R. A.: Evidence for the identity of the NADH phosphate specific sulphite and nitrite reductases of E. coli. J. biol. Chem. 238, 3466–3471 (1963).
Kessler, E.: Nitrate assimilation by plants. Ann. Rev. Plant Physiol. 15, 57–72 (1964).
Kessler, E., Czygan, F. C.: Seasonal changes in the nitrate-reducing activity of a green algae (Ankistrodesmus braunii). Experientia (Basel) 19, 89–90 (1963).
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., Randall, R. S.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951).
Mayer, A. M.: Subcellular location of sulphite reductase in plant tissues. Plant Physiol. 42, 324–326 (1967).
Miflin, B. J.: Distribution of nitrate and nitrite reductase in barley. Nature (Lond.) 214, 1133–1134 (1967).
Miflin, B. J.: Nitrate and nitrite reductase systems in barley roots. Rev. roum. Biochim. 7, 53–60 (1970a).
Miflin, B. J.: Studies on the subcellular location of particulate nitrate and nitrite reductase, glutamic dehydrogenase and other enzymes in barley roots. Planta (Berl.) 93, 160–170 (1970b).
Paneque, A., Ramirez, J. M., Del Campo, F. F., Losada, M.: Light and dark reduction of nitrite in a reconstituted enzyme system. J. biol. Chem. 239, 1737–1741 (1964).
Russel, J. A.: The estimation of small amounts of ammonia by the phenol-hypochlorite reaction. J. biol. Chem. 156, 457–465 (1944).
Siegel, L. M.: A direct microdetermination for sulphide. Analyt. Biochem. 11, 126–132 (1965).
Tagawa, K., Arnon, D. I.: Ferredoxins as electron carriers in photosynthesis and in the biological production of hydrogen gas. Nature (Lond.) 195, 537–543 (1962).
Wallace, W., Pate, J. S.: Nitrate reductase in the field pea. Ann. Botany 29, 655 (1965).
Wallace, W., Pate, J. S.: Nitrate assimilation in hihher plants with special reference to the Cockleburr. Ann. Botany 31, 213–228 (1967).
Wessels, J. S. C.: DNP as a catalyst of photosynthetic phosphorylation. Biochem. biophys. Acta (Amst.) 36, 264–265 (1959).
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Bourne, W.F., Miflin, B.J. Studies on nitrite reductase in barley. Planta 111, 47–56 (1973). https://doi.org/10.1007/BF00386734
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DOI: https://doi.org/10.1007/BF00386734