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Nitrite inhibition of nitrogenase from soybean bacteroids

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Abstract

Nitrogenase from soybean bacteroids was purified and used to study NO −2 effects either as unfractionated enzyme or as reconstituted enzyme from separated nitrogenase components I and II. Partially purified enzyme was strongly inhibited by nitrite at concentrations less than 0.1 mM. This inhibition was typically referred to as competitive with an inhibition constant (K i) for NO −2 which was 5.2 mM. Kinetics studies showed an abnormally low apparent constant of association between enzyme and NO −2 (k a=60 M-1·s-1). Nitrite appeared to bind to the MoFe protein, without any effect on Fe component, giving a completely reversible inhibition. Nitrite was found not to be an alternative substrate for nitrogenase.

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Abbreviations

TES:

N-tris (hydroxymethyl) methyl-2-aminoethane sulfonic acid

PPG:

Polypropylene glycol

References

  • Apte, S. K., David, K. A. V., Thomas, J.: Conformational changes in the nitrogenase complex in vivo by preincubation under acetylene. Biochem. Biophys. Res. Commun. 83, 1157–1163 (1978)

    PubMed  Google Scholar 

  • Bergersen, F. J., Turner, G. L.: Kinetic studies of nitrogenase from soya-bean root-nodule bacteroids. Biochem. J. 131, 61–75 (1973)

    PubMed  Google Scholar 

  • Burns, R. C., Fuchsman, W. H., Hardy, R. W. F.: Nitrogenase from vanadium-grown Azotobacter: Isolation, characteristics, and mechanistic implications. Biochem. Biophys. Res. Commun. 42, 353–358 (1971)

    PubMed  Google Scholar 

  • Gibson, A. H.: Recovery and compensation by nodulated legumes to environmental stress. In: Symbiotic nitrogen fixation in plants (P. S. Nutman, ed.), pp. 385–403. Cambridge: Cambridge University Press 1976

    Google Scholar 

  • Gibson, A. H., Pagan, J. D.: Nitrate effects on the nodulation of legumes inoculated with nitrate-reductase-deficient mutants of Rhizobium. Planta 134, 17–22 (1977)

    Google Scholar 

  • Hardy, R. W. F., Holsten, R. D., Jackson, E. K., Burns, R. C.: The acetylene-ethylene assay for N2 fixation: Laboratory and field evaluation. Plant Physiol. 43, 1185–1207 (1968)

    Google Scholar 

  • Hwang, J. C., Chen, C. H., Burris, R. H.: Inhibition of nitrogenasecatalyzed reductions. Biochim. Biophys. Acta 292, 256–270 (1973)

    PubMed  Google Scholar 

  • Israel, D. W., Howard, R. L., Evans, H. J., Russel, S. A.: Purification and characterization of the molybdenum-iron protein component of nitrogenase from soybean nodule bacteroids. J. Biol. Chem. 249, 500–508 (1974)

    PubMed  Google Scholar 

  • Kamberger, W.: Regulation of symbiotic nitrogen fixation in root nodules of Alfalfa (Medicago sativa) infected with Rhizobium meliloti. Arch. Microbiol. 115, 103–108 (1977)

    PubMed  Google Scholar 

  • Kennedy, I. R., Rigaud, J., Trinchant, J. C.: Nitrate reductase from bacteroids of Rhizobium japonicum: Enzyme characteristics and possible interactions with nitrogen fixation. Biochim. Biophys. Acta 397, 24–35 (1975)

    PubMed  Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)

    PubMed  Google Scholar 

  • Nicholas, D. J. D., Nason, A.: Determination of nitrate and nitrite. In: Methods in enzymology, Vol. III (S. P. Colowick, N. O. Kaplan, eds.), pp. 981–984. London: Academic Press 1957

    Google Scholar 

  • Rigaud, J.: Effet des nitrates sur la fixation d'azote par les nodules de haricot (Phaseolus vulgaris L.). Physiol. vég. 14, 297–308 (1976)

    Google Scholar 

  • Rigaud, J., Bergersen, F. J., Turner, G. L., Daniel, R. M.: Nitrate dependent anaerobic acetylene-reduction and nitrogen fixation by soybean bacteroids. J. gen. Microbiol. 77, 137–144 (1973)

    Google Scholar 

  • Rigaud, J., Puppo, A.: Indol-3-acetic acid catabolism by soybean bacteroids. J. gen. Microbiol. 88, 223–228 (1975)

    Google Scholar 

  • Rigaud, J., Puppo, A.: Effect of nitrite upon leghemoglobin and interaction with nitrogen fixation. Biochim. Biophys. Acta 497, 702–706 (1977)

    PubMed  Google Scholar 

  • Rivera-Ortiz, J. M., Burris, R. H.: Interactions among substrates and inhibitors of nitrogenase. J. Bacteriol. 123, 537–545 (1975)

    PubMed  Google Scholar 

  • Schöllhorn, R., Burris, R. H.: Acetylene as a competitive inhibitor of N2 fixation. Proc. Natl. Acad. Sci. U.S.A. 58, 213–216 (1967)

    PubMed  Google Scholar 

  • Seto, B. L., Mortenson, L. E.: Mechanism of carbamyl phosphate inhibition of nitrogenase of Clostridium pasteurianum. J. Bacteriol. 117, 805–812 (1974)

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Faculté des Sciences et des Techniques, Laboratoire de Biologie végétale, Parc Valrose, 06034, Nice Cedex, France

    J. C. Trinchant & J. Rigaud

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  1. J. C. Trinchant
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  2. J. Rigaud
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Trinchant, J.C., Rigaud, J. Nitrite inhibition of nitrogenase from soybean bacteroids. Arch. Microbiol. 124, 49–54 (1980). https://doi.org/10.1007/BF00407027

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  • DOI: https://doi.org/10.1007/BF00407027

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