Archives of Microbiology

, Volume 126, Issue 2, pp 155–159 | Cite as

Effects of molybdenum and tungsten on induction of nitrate reductase and formate dehydrogenase in wild type and mutant Paracoccus denitrificans

  • Kathleen A. Burke
  • Kathleen Calder
  • June Lascelles
Article

Abstract

Molybdenum is required for induction of nitrate reductase and of NAD-linked formate dehydrogenase activities in suspensions of wild type Paracoccus denitrificans; tungsten prevents the development of these enzyme activities. The wild type forms a membrane protein Mr150,000 when incubated with tungsten and inducers of nitrate reductase and this is presumed to represent an inactive form of the enzyme. Suspensions of mutant M-1 did not develop nitrate reductase or formate dehydrogenase activities but the membrane protein Mr150,000 was formed under all conditions tested, including without inducers and without molybdenum. Analysis of membranes, solubilized with deoxycholate, by polyacrylamide gel electrophoresis under nondenaturing conditions showed that the mutant protein had similar electrophoretic mobility to the active nitrate reductase formed by the wilde type. Autoradiography of preparations from cells incubated with 55Fe showed that the mutant and wild type proteins contained iron. However, in similar experiments with 99Mo, incorporation of molybdenum into the mutant protein was not detectable.

We conclude that mutant M-1 is defective in one or more steps required to process molybdenum for incorporation into molybdoenzymes. This failure affects the normal regulation of nitrate reductase protein with respect to the role of inducers.

Key words

Nitrate reductase Formate dehydrogenase Molybdenum Tungsten Paracoccus dentrificans Inducers Membrane proteins 

Non-Standard Abbreviations

DOC

deoxycholate

PAGE

polyacrylamide gel electrophoresis

SDS

sodium dodecyl sulfate

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Bamforth, C. W., Quayle, J. R.: Aerobic and anaerobic growth of paracoccus denitrificans on methanol. Arch. Microbiol. 119, 91–97 (1978)Google Scholar
  2. Bonner, W. M., Laskey, R. A.: A film detection method for tritiumlabelled protein and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88 (1974)Google Scholar
  3. Burke, K. A., Lascelles, J.: Nitrate reductase system in Staphylococcus aureus wild type and mutants. J. Bacteriol. 123, 308–316 (1975)Google Scholar
  4. Calder, K., Burke, K.A., Lascelles, J.: Induction of nitrate reductase and membrane cytochromes in wild type and chlorate-resistant Paracoccus denitrificans (Submitted)Google Scholar
  5. Cox, R. B., Quayle, J. R.: The autotrophic growth of Micrococcus denitrificans on methanol. Biochem. J. 150, 569–571 (1975)Google Scholar
  6. Dulley, J. R., Greene, P. A.: A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal. Biochem. 64, 136–141 (1975)Google Scholar
  7. Enoch, H. G., Lester, R. L.: The purification and properties of formate dehydrogenase and nitrate reductase from Escherichia coli. J. Biol. Chem. 250, 6693–6705 (1975)Google Scholar
  8. Glaser, J. H., DeMoss, J. A.: Comparison of nitrate reductase mutants of Escherichia coli selected by alternative procedures. Mol. Gen. Genet. 116, 1–10 (1972)Google Scholar
  9. Johnson, P. A., Quayle, J. R.: Microbial growth on C1 compounds. 6. Oxidation of methanol, formaldehyde and formate by methanolgrown Pseudomonas AM1. Biochem. J. 93, 281–290 (1964)Google Scholar
  10. Lascelles, J., Burke, K. A.: Reduction of ferric iron by l-lactate and DL-glycerol-3-phosphate in membrane preparations from Staphylococcus aureus and interactions with the nitrate reductase system. J. Bacteriol. 134, 585–589 (1978)Google Scholar
  11. MacGregor, C. H.: Synthesis of nitrate reductase components in chlorate resistant mutants of Escherichia coli. J. Bacteriol. 121, 1117–1121 (1975)Google Scholar
  12. Pienkos, P. T., Shah, V. K., Brill, W. J.: Molybdenum cofactors from molybdoenzymes and in vitro reconstitution of nitrogenase and nitrate reductase. Proc. Natl. Acad. Sci. USA 74, 5468–5471 (1977)Google Scholar
  13. Scott, R. H., DeMoss, J. A.: Formation of the formate-nitrate electron transport pathway from inactive components in Escherichia coli. J. Bacteriol. 126, 478–486 (1976)Google Scholar
  14. Scott, R. H., Sperl, G. T., DeMoss, J. A.: In vitro incorporation of molybdate into demolybdoproteins in Escherichia coli. J. Bacteriol. 137, 719–726 (1979)Google Scholar
  15. Shah, V. K., Brill, W. J.: Isolation of an iron-molybdenum cofactor from nitrogenase. Proc. Natl. Acad. Sci. USA 34, 3249–3253 (1977)Google Scholar
  16. Sperl, G. T., DeMoss, J. A.: chl D gene function in molydate activation of nitrate reductase. J. Bacteriol. 122, 1230–1238 (1975)Google Scholar
  17. Stouthamer, A. H.: Biochemistry and genetics of nitrate reductase in bacteria. Adv. Microbial Physiol. 14, 315–375 (1976)Google Scholar
  18. Thauer, R. K., Fuchs, G., Jungermann, K.: Role of iron-sulfur proteins in formate metabolism. In: Iron-sulfur Proteins, Vol. III (W. Lovenberg, ed.). pp. 121–156 New York: Academic Press 1977Google Scholar

Copyright information

© Springer-Verlag 1980

Authors and Affiliations

  • Kathleen A. Burke
    • 1
  • Kathleen Calder
    • 1
  • June Lascelles
    • 1
  1. 1.Department of MicrobiologyUniversity of CaliforniaLos AngelesUSA

Personalised recommendations