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. 1980 May;65(5):944–948. doi: 10.1104/pp.65.5.944

Synthesis of Nitrate Reductase in Chlorella

II. EVIDENCE FOR SYNTHESIS IN AMMONIA-GROWN CELLS 1

Edward A Funkhouser 1,2, Candadai S Ramadoss 1,2
PMCID: PMC440453  PMID: 16661311

Abstract

Antiserum was prepared against nitrate reductase (EC 1.6.6.1) purified to homogeneity from Chlorella vulgaris Beijerinck. Both crude antiserum and anti-nitrate reductase antibodies prepared from it were used as re-agents to study the synthesis of nitrate reductase. Cell extracts from cultures which were grown with ammonia salts as the sole source of nitrogen contained almost no active enzyme. These extracts did contain material which binds to antibody and is thus immunologically related to purified nitrate reductase. The presence of this cross reacting material in cell extracts was detected by the ability of these extracts to (a) lower the titer of antisera; (b) form a biphasic precipitin curve with purified antibody; and (c) increase the peak height of a standard amount of purified nitrate reductase in rocket immunoelectrophoresis assay. These results suggest that ammonia-grown cells contain nitrate reductase precursor protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amy N. K., Garrett R. H. Immunoelectrophoretic determination of nitrate reductase in Neurospora crassa. Anal Biochem. 1979 May;95(1):97–107. doi: 10.1016/0003-2697(79)90191-x. [DOI] [PubMed] [Google Scholar]
  2. Funkhouser E. A. Synthesis of Nitrate Reductase in Chlorella: I. EVIDENCE FOR AN INACTIVE PROTEIN PRECURSOR. Plant Physiol. 1980 May;65(5):939–943. doi: 10.1104/pp.65.5.939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Giri L., Ramadoss C. S. Physical studies on assimilatory nitrate reductase from Chlorella vulgaris. J Biol Chem. 1979 Nov 25;254(22):11703–11712. [PubMed] [Google Scholar]
  4. Jackson A. O., Larkins B. A. Influence of Ionic Strength, pH, and Chelation of Divalent Metals on Isolation of Polyribosomes from Tobacco Leaves. Plant Physiol. 1976 Jan;57(1):5–10. doi: 10.1104/pp.57.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jetschmann K., Solomonson L. P., Vennesland B. Activation of nitrate reductase by oxidation. Biochim Biophys Acta. 1972 Aug 17;275(2):276–278. doi: 10.1016/0005-2728(72)90048-5. [DOI] [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Losada M., Paneque A., Aparicio P. J., Vega J. M., Cárdenas J., Herrera J. Inactivation and repression by ammonium of the nitrate reducing system in chlorella. Biochem Biophys Res Commun. 1970 Mar 27;38(6):1009–1015. doi: 10.1016/0006-291x(70)90340-2. [DOI] [PubMed] [Google Scholar]
  8. Nicholas D. J., Ferrante J. V., Clarke G. R. Immunological studies with nitrogenase from Azotobacter and bacterial nitrate reductases. Anal Biochem. 1979 May;95(1):24–31. doi: 10.1016/0003-2697(79)90180-5. [DOI] [PubMed] [Google Scholar]
  9. Notton B. A., Graf L., Hewitt E. J., Povey R. C. The role of molybdenum in the synthesis of nitrate reductase in cauliflower (Brassica oleracea L. var Botrytis L.) and spinach (Spinacea oleracea L.). Biochim Biophys Acta. 1974 Sep 11;364(1):45–58. doi: 10.1016/0005-2744(74)90131-4. [DOI] [PubMed] [Google Scholar]
  10. Premakumar R., Sorger G. J., Gooden D. Stability of messenger RNA for nitrate reductase in Neurospora crassa. Biochim Biophys Acta. 1978 Jun 22;519(1):275–278. doi: 10.1016/0005-2787(78)90080-1. [DOI] [PubMed] [Google Scholar]
  11. Solomonson L. P., Lorimer G. H., Hall R. L., Borchers R., Bailey J. L. Reduced nicotinamide adenine dinucleotide-nitrate reductase of Chlorella vulgaris. Purification, prosthetic groups, and molecular properties. J Biol Chem. 1975 Jun 10;250(11):4120–4127. [PubMed] [Google Scholar]
  12. Solomonson L. P. Purification of NADH-Nitrate Reductase by Affinity Chromatography. Plant Physiol. 1975 Dec;56(6):853–855. doi: 10.1104/pp.56.6.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Solomonson L. P., Vennesland B. Nitrate Reductase and Chlorate Toxicity in Chlorella vulgaris Beijerinck. Plant Physiol. 1972 Oct;50(4):421–424. doi: 10.1104/pp.50.4.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Solomonson L. P., Vennesland B. Properties of a nitrate reductase of Chlorella. Biochim Biophys Acta. 1972 Jun 23;267(3):544–557. doi: 10.1016/0005-2728(72)90183-1. [DOI] [PubMed] [Google Scholar]
  15. Vennesland B., Jetschmann C. The nitrate reductase of Chlorella pyrenoidosa. Biochim Biophys Acta. 1971 Mar 10;227(3):554–564. doi: 10.1016/0005-2744(71)90006-4. [DOI] [PubMed] [Google Scholar]
  16. van 't Riet J., Wientjes F. B., van Doorn J., Planta R. J. Purification and characterization of the respiratory nitrate reductase of Bacillus licheniformis. Biochim Biophys Acta. 1979 Feb 26;576(2):347–360. doi: 10.1016/0005-2795(79)90410-0. [DOI] [PubMed] [Google Scholar]

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