Skip to main content
Log in

Periplasmic location of nitrous oxide reductase and its apoform in denitrifying Pseudomonas stutzeri

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Immunogold labelling techniques on ultrathin sections of low temperature embedded cells yielded evidence for the periplasmic location of the respiratory enzymes N2O reductase and nitrite reductase (cytochrome cd 1) in Pseudomonas stutzeri strain ZoBell. Cell fractionation by spheroplast preparation and two-dimensional electrophoresis showed the absence of a membrane association of these enzymes. Immunocytochemical localization of N2O reductase in a mutant strain deficient in the chromophore of N2O reductase showed the gold label at the cell periphery, indicating that the copper chromophore processing takes place after export of this protein's apoform.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Coyne MS, Arunakumari A, Pankratz HS, Tiedje JM (1990) Localization of the cytochrome cd 1 and copper nitrite reductase in denitrifying bacteria. J Bacteriol 172: 2558–2562

    Google Scholar 

  • Blümle S, Zumft WG (1991) Respiratory nitrate reductase from denitrifying Pseudomonas stutzeri, purification, properties and target of proteolysis. Biochim Biophys Acta 1057: 102–108

    Google Scholar 

  • Dermastia M, Turk T, Hollocher T (1991) Nitric oxide reductase purification from Paracoccus denitrificans with use of a single column and some characteristics. J Biol Chem 266: 10899–10905

    Google Scholar 

  • Ferguson SJ (1987) Denitrification: a question of the control and organization of electron and ion transport. Trends Biochem Sci 12: 354–357

    Google Scholar 

  • Görg A, Postel W, Westermeier R, Gianazza E, Righetti PG (1980) Gel gradient electrophoresis, isoelectric focusing, and two-dimensional techniques in horizontal, ultrathin polyacrylamide layers. J Biochem Biophys Methods 3: 273–284

    Google Scholar 

  • Heiss B, Frunzke K, Zumft WG (1989) Formation of the N-N bond from nitric oxide by a membrane-bound cytochrome bc complex of nitrate-respiring (denitrifying) Pseudomonas stutzeri. J Bacteriol 171: 3288–3297

    Google Scholar 

  • Hochstein LI, Tomlinson GA (1989) The enzymes associated with denitrification. Annu Rev Microbiol 41: 231–261

    Google Scholar 

  • Jones RW, Lamont A, Garland PB (1978) The mechanism of protein translocation driven by the respiratory nitrate reductase complex of Escherichia coli. Biochem J 190: 79–94

    Google Scholar 

  • Jüngst A, Wakabayashi S, Matsubara H, Zumft WG (1991) The nir STBM region coding for cytochrome cd 1-dependent nitrite respiration Pseudomonas stutzeri consists of a cluster of mono-, di-, and tetraheme proteins. FEBS Lett 279: 205–209

    Google Scholar 

  • Körner H, Zumft WG (1989) Expression of denitrification enzymes in response to the dissolved oxygen level and respiratory substrate in continuous culture of Pseudomonas stutzeri. Appl Environ Microbiol 55: 1670–1676

    Google Scholar 

  • Mancini G, Carbonara AO, Heremans JF (1965) Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry 2: 235–238

    Google Scholar 

  • McEwan AG, Ferguson SJ, Jackson JB, Meyer O, Wetzstein HG (1987) The periplasmic nitrate reductase of Rhodobacter capsulatus — Purification, characterization and distinction from a single reductase for trimethylamine-N-oxide, dimethylsulfoxide and chlorate. Arch Microbiol 147: 340–345

    Google Scholar 

  • Minagawa N, Zumft WG (1988) Cadium-copper antagonism in the activation of periplasmic nitrous oxide reductase of copperdeficient cells from Pseudomonas stutzeri. Biol Metals 1: 117–121

    Google Scholar 

  • Mokhele K, Tang YJ, Clark MA, Ingraham JL (1987) A Pseudomonas stutzeri outer membrane protein inserts copper into N2O reductase. J Bacteriol 169: 5721–5726

    Google Scholar 

  • O'Farrell PH (1975) High-resolution two-dimensional electrophoresis of proteins. J Biol Chem 250: 4007–4021

    Google Scholar 

  • Pages JM, Anba J, Bernadac A, Shinagawa H, Nakata A, Lazdunski C (1984) Normal precursors of periplasmic proteins accumulated in the cytoplasm are not exported post-translationally in Escherichia coli. Eur J Biochem 143: 499–505

    Google Scholar 

  • Rohde M, Gerberding H, Mund T, Kohring GW (1988) Immunoelectron microscopic localization of bacterial enzymes: pre-and post-embedding labelling techniques on resin-embedded samples In: Mayer F (ed) Methods in microbiology, vol 20. Academic Press, London, pp 175–210

    Google Scholar 

  • Sammons DW, Adams LD, Nishizawa EE (1981) Ultrasensitive silver-based color staining of polypeptides in polyacrylamide gels. Electrophoresis 2: 135–141

    Google Scholar 

  • Saraste M, Kuronen T (1978) Interaction of Pseudomonas cytochrome cd 1 with the cytoplasmic membrane. Biochim Biophys Acta 513: 117–131

    Google Scholar 

  • Viebrock A, Zumft WG (1988) Molecular cloning, heterologous expression, and primary structure of the structural gene for the copper enzyme nitrous oxide reductase from denitrifying Pseudomonas stutzeri. J Bacteriol 170: 4658–4668

    Google Scholar 

  • vanVerseveld HW, Bosma G (1987) The respiratory chain and energy conservation in the mitochondrion-like bacterium Paracoccus denitrificans. Microbiol Sci 4: 329–333

    Google Scholar 

  • Wood PM (1978) Periplasmic location of the terminal reductase in nitrite respiration. FEBS Lett 92: 214–218

    Google Scholar 

  • Zumft WG, Döhler K, Körner H (1985) Isolation and characterization of transposon Tn5-induced mutants of Pseudomonas perfectomarina defective in nitrous oxide respiration. J Bacteriol 163: 918–924

    Google Scholar 

  • Zumft WG, Döhler K, Körner H, Löchelt S, Viebrock A, Frunzke K (1988) Defects in cytochrome cd 1-dependent nitrite respiration of transposon Tn5-induced mutants from Pseudomonas stutzeri. Arch Microbiol 149: 492–498

    Google Scholar 

  • Zumft WG, Viebrock-Sambale A, Braun C (1990) Nitrous oxide reductase from denitrifying Pseudomonas stutzeri. Genes for copper-processing and properties of the deduced products, including a new member of the family of ATP/GTP-binding proteins. Eur J Biochem 192: 591–599

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Körner, H., Mayer, F. Periplasmic location of nitrous oxide reductase and its apoform in denitrifying Pseudomonas stutzeri . Arch. Microbiol. 157, 218–222 (1992). https://doi.org/10.1007/BF00245153

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00245153

Key words

Navigation