Characterization of the periplasmic cytochromes of : identification of the electron acceptor for methanol dehydrogenase, and description of a novel cytochrome heterodimer Free

Abstract

Summary

This paper describes periplasmic -type cytochromes from two strains of NCIB 8944 grown in heterotrophic or methylotrophic conditions. It is shown that the functions of two monomeric, monohaem cytochromes induced during growth on methanol have been wrongly designated in previous work. The CO-reactive cytochrome (30 kDa) is not the electron acceptor for methanol dehydrogenase; this is shown to be the role of the cytochrome (22 kDa). The monomeric 45 kDa cytochrome induced in conditions of oxygen insufficiency is a dihaem -type cytochrome and does not contain haem as previously assumed. In addition to these cytochromes, the Oxford strain of NCIB 8944 contains two cytochrome complexes. One of these (150 kDa), produced in relatively small amounts, consists of a non-haem protein plus four haemoproteins (28, 33, 41 and 47 kDa). The second complex is a novel dimeric multi-haem cytochrome (46 kDa) which constitutes about 25% of the periplasmic -type cytochrome. It reacts with CO and has no methionine ligands. One subunit (16 kDa) has two low-spin haems; the larger subunit (30 kDa) has three haems which have low-spin characteristics in the oxidized state and are high-spin in the reduced state. The subunits were readily separated at pH 12 and could be subsequently reconstituted into a complex indistinguishable from the original. The 30 kDa subunit was denatured on prolonged exposure to high pH, which also converted it to a low-spin cytochrome. No function could be designated for these novel -type cytochrome complexes.

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1991-02-01
2024-03-28
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References

  1. Alefounder P. R., Ferguson S. J. 1981; A periplasmic location for methanol dehydrogenase from Paracoccus denitrificans: implications for proton pumping by cytochrome aa 3 . Biochemical and Biophysical Research Communications 98:778–784
    [Google Scholar]
  2. Anthony C. 1982 The Biochemistry of Methylotrophs. London: Academic Press;
    [Google Scholar]
  3. Anthony C. 1986; The bacterial oxidation of methane and methanol. Advances in Microbial Physiology 27:113–210
    [Google Scholar]
  4. Anthony C. 1988; Quinoproteins and energy transduction. Bacterial Energy Transduction293–316 Anthony C. London: Academic Press;
    [Google Scholar]
  5. Anthony C. 1989; Quinoproteins in C-l dissimilation by bacteria. Antonie van Leeuwenhoek 56:13–23
    [Google Scholar]
  6. Anthony C., Zatman L. J. 1967; The microbial oxidation of methanol: purification and properties of the alcohol dehydrogenase of Pseudomonas sp. M27. Biochemical Journal 104:953–959
    [Google Scholar]
  7. Beardmore-Gray M, O’Keeffe D. T., Anthony C. 1983; The methanol: cytochrome c oxidoreductase activity of methylotrophs.. Journal of General Microbiology 129:923–933
    [Google Scholar]
  8. Berry E. A., Trumpower B. L. 1985; Isolation of ubiquinol oxidase from Paracoccus denitrificans and resolution into cytochrome bc 1 and cytochrome c-aa 3 complexes.. Journal of Biological Chemistry 260:2458–2467
    [Google Scholar]
  9. Berry M. J., George S. J., Thomson A. J., Santos H., Turner D. L. 1990; Cytochrome c" isolated from Methylophilus methylotro-phus. An example of bis-histidine-co-ordinated Fe3+ haem, with near-perpendicular orientation of the ligands.. Biochemical Journal 270:413–417
    [Google Scholar]
  10. Bosma G., Braster M., Stouthamer A. H., van Verseveld H. W. 1987a; Isolation and characterization of ubiquinol oxidase complexes from Paracoccus denitrificans cells cultured under various limiting growth conditions in the chemostat. European Journal of Biochemistry 165:657–665
    [Google Scholar]
  11. Bosma G., Braster M., Stouthamer A. H., van Verseveld H. W. 1987b; Subfractionation and characterization of soluble c-type cytochromes from Paracoccus denitrificans cultured under various limiting conditions in the chemostat. European Journal of Biochemistry 165:665–670
    [Google Scholar]
  12. Bradford M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
    [Google Scholar]
  13. Davidson V. L., Kumar M. A. 1989; Cytochrome c-550 mediates electron transfer from inducible periplasmic c-type cytochromes to the cytoplasmic membrane of Paracoccus denitrificans . FEBS tetters 245:271–273
    [Google Scholar]
  14. Day D. J., Anthony C. 1990; Methanol dehydrogenase from Methylobacterium extorquens AMI.. Methods in Enzymology 188:210–216
    [Google Scholar]
  15. Day D. J., Nunn D. N., Anthony C. 1990; Characterization of a novel soluble c-type cytochrome in a moxD mutant of Methylobacterium extorquens AMI.. Journal of General Microbiology 136:181–188
    [Google Scholar]
  16. Dutton P. L. 1978; Redox potentiometry: determination of midpoint potentials of oxidation-reduction components of biological electron transfer systems. Methods in Enzymology 54:411–434
    [Google Scholar]
  17. Ferguson S. J. 1988; Periplasmic electron transport reactions. Bacterial Energy Transduction141–182 Edited by Anthony C. London: Academic Press;
    [Google Scholar]
  18. Froud S. J., Anthony C. 1984; The roles of cytochrome c in membranes of Methylophilus methylotrophus . Journal of General Microbiology 130:3319–3325
    [Google Scholar]
  19. Fuhrhop J. H., Smith K. M. 1975; Laboratory methods. Porphyrins and Metalloporphyrins757–869 Edited by Smith K. M. Amsterdam: Elsevier;
    [Google Scholar]
  20. Gray K. A., Knaff D. B., Husain M., Davidson V. L. 1986; Measurement of the oxidation-reduction potentials of amicyanin and c-type cytochromes from Paracoccus denitrificans . FEBS Letters 207:239–242
    [Google Scholar]
  21. Harms N, de Vries G., Maurer K., Hoogendijk J., Stouthamer A. H. 1987; Isolation and nucleotide sequence of the methanol dehydrogenase structural gene from Paracoccus denitrificans . Journal of Bacteriology 169:3969–3975
    [Google Scholar]
  22. Husain M., Davidson V. L. 1986; Characterization of two inducible periplasmic c-type cytochromes from Paracoccus denitrificans . Journal of Biological Chemistry 261:8577–8580
    [Google Scholar]
  23. Kornberg H. L., Morris J. G. 1965; The utilization of glycollate by Micrococcus denitrificans: the beta hydroxyaspartate pathway.. Biochemical Journal 95:557–586
    [Google Scholar]
  24. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature London: 227680–685
    [Google Scholar]
  25. Lambin P., Rochu D., Fine J. M. 1976; A new method for determination of molecular weights of proteins by electrophoresis across a sodium dodecyl sulphate polyacrylamide gradient gel. Analytical Biochemistry 74:567–575
    [Google Scholar]
  26. Nunn D. N., Anthony C. 1988; The nucleotide sequence and deduced amino acid sequence of the cytochrome cL gene of Methylobacterium extorquens AMI: a novel class of c-type cytochromes.. Biochemical Journal 256:673–676
    [Google Scholar]
  27. Page M. D., Ferguson S. J. 1989; A bacterial c-type cytochrome can be translocated to the periplasm as an apo form; the biosynthesis of cytochrome cd 1 (nitrate reductase) from Paracoccus denitrificans . Molecular Microbiology 3:653–631
    [Google Scholar]
  28. Page M. D., Ferguson S. J. 1990; Apo-forms of cytochrome c-550 and cytochrome a/, are translocated to the periplasm of Paracoccus denitrificans in the absence of haem incorporation caused by either mutation or inhibition of haem synthesis.. Molecular Microbiology 4:1181–1192
    [Google Scholar]
  29. Raitio M., Pispa J. M., Metso T, Saraste M. 1990; Are there isoenzymes of cytochrome c oxidase in Paracoccus denitrificans?. FEBS Letters 261:431–435
    [Google Scholar]
  30. Santos H., Turner D. L. 1988; Characterization and NMR studies of a novel cytochrome c isolated from Methylophilus methylotrophus which shows a redox-linked change of spin state.. Biochimica et Biophysica Acta 954:277–286
    [Google Scholar]
  31. Smith A., Hill S., Anthony C. 1988; A haemoprotein is not involved in the control by oxygen of enteric nitrogenase synthesis. Journal of General Microbiology 134:1499–1507
    [Google Scholar]
  32. van Spanning R. J. M., Wansell C, Harms N., Oltmann L. F., Stouthamer A. H. 1990; Mutagenesis of the gene encoding cytochrome c-550 of Paracoccus denitrificans and analysis of the resultant physiological effects.. Journal of Bacteriology 172:986–996
    [Google Scholar]
  33. Thomas P. E., Ryan D., Levin W. 1986; An improved staining procedure for the detection of the peroxidase activity of cytochrome P450 on SDS-polyacrylamide gels. Analytical Biochemistry 75:168–176
    [Google Scholar]
  34. van Verseveld H. W., Stouthamer A. H. 1978; Electron transport chain and coupled oxidative phosphorylation in methanol-grown Paracoccus denitrificans . Archives of Microbiology 118:13–20
    [Google Scholar]
  35. de Vries G. E. 1986; Molecular biology of bacterial methanol oxidation. FEMS Microbiology Reviews 39:235–258
    [Google Scholar]
  36. De Vries G. E., Kues U., Stahl U. 1990; Physiology and genetics of methylotrophic bacteria. FEMS Microbiology Reviews 75:57–102
    [Google Scholar]
  37. Weber K., Osborn M. 1975; Proteins and sodium dodecyl sulphate: molecular weight determination on polyacrylamide gels and related procedures. The Proteins, 3rd edn. 1179–223 Neurath H, Hill R. L. London: Academic Press;
    [Google Scholar]
  38. Wood P. M. 1984; Bacterial proteins with CO-binding b- or c-type haem. Functions and absorption spectroscopy. Biochimica et Biophysica Acta 768:293–317
    [Google Scholar]
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