Skip to main content
SpringerLink
Log in

Cytochromes c of the anaerobic methacrylate reducer Geobacter sulfurreducens AM-1

  • Experimental Articles
  • Published:
Microbiology Aims and scope Submit manuscript

Abstract

Cytochromes c were found in the cells of the bacterium Geobacter sulfurreducens AM-1 grown on acetate and methacrylate. The periplasmic extract of G. sulfurreducens AM-1 contained about 88% of the total content of cytochromes c of intact cells. The analysis of cytochromes c from the native cells of G. sulfurreducens AM-1, from the periplasmic extract and from the cells treated by an alkaline solution showed the presence of nine proteins containing heme c. The molecular masses of cytochromes c from G. sulfurreducens AM-1 were 12.5, 15.5, 25.7, 29.5, 34.7, 41.7, 50.1, 63.1, and 67.6 kDa; localization of each cytochrome c was determined. Three heme-containing proteins (15.5 kDa, 25.7 kDa, and 29.5 kDa with the most intensive staining) were present mainly in the periplasm of the bacterium. The other two (50.1 and 67.6 kDa) were supposedly localized in the cell membrane. Cytochromes c with the molecular masses of 12.5, 15.5, and 67.6 kDa are considered as possible components of the methacrylate redox system of G. sulfurreducens AM-1.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Caccavo, F.J.R., Lonergan, D.J., Lovley, D.R., Davis, M., Stolz, J.F., and McInerney, M.J., Geobacter sulfurreducens sp. nov., a Hydrogen- and Acetate-Oxidizing Dissimilatory Metal-Reducing Microorganism, Appl. Environ. Microbiol., 1994, vol. 60, no. 10, pp. 3752–3759.

    PubMed  CAS  Google Scholar 

  2. Kaufmann, F. and Lovley, D.R., Isolation and Characterization of a Soluble NADPH-Dependent Fe(III) Reductase from Geobacter sulfurreducens, J. Bacteriol., 2001, vol. 183, no. 15, pp. 4468–4476.

    Article  PubMed  CAS  Google Scholar 

  3. Methe, B.A., Nelson, K.E., Eisen, J.A., Paulsen, I.T., Nelson, W., Heidelberg, J.F., Wu, D., Wu, M., Ward, N., Beanan, M.J., Dodson, R.J., Madupu, R., Brinkac, L.M., Daugherty, S.C., DeBoy, R.T., Durkon, A.S., Gwinn, M., Kolonay, J.F., Sullivan, S.A., Haft, D.H., Selengut, J., Davidsen, T.M., Zafar, N., White, O., Tran, B., Romero, C., Forberger, H.A., Weidman, J., Khouri, H., Feldblyum, T.V., Utterback, T.R., Van Aken, S.E., Lovley, D.R., and Frazer, C.M., Genome of Geobacter sulfurreducens: Metal Reduction in Subsurface Environments, Science, 2003, vol. 302, pp. 1967–1969.

    Article  PubMed  CAS  Google Scholar 

  4. Butler, J.E., Kaufmann, F., Coppi, M.V., Nunez, C., and Lovley, D.R., MacA, a Diheme c-Type Cytochrome Involved in Fe-(III) Reduction by Geobacter sulfurreducens, J. Bacteriol., 2004, vol. 186, pp. 4042–4045.

    Article  PubMed  CAS  Google Scholar 

  5. Leang, C., Coppi, M.V., and Lovley, D.R., OmcB, a c-Type Polyheme Cytochrome, Involved in Fe(III) Reduction in Geobacter sulfurreducens, J. Bacteriol., 2003, vol. 185, pp. 2096–2103.

    Article  PubMed  CAS  Google Scholar 

  6. Leang, C., Adams, L.A., Chin, K.-J., Nevin, K.P., Methé, B.A., Webster, J., Sharma, M.L., and Lovley, D.R., Adaptation To Disruption of the Electron Transfer Pathway for Fe(III) Reduction in Geobacter sulfurreducens, J. Bacteriol., 2005, vol. 187, no. 17, pp. 5918–5926.

    Article  PubMed  CAS  Google Scholar 

  7. Kim, B.C., Qian, X., Leang, C., Coppi, N.V., and Lovley, D.R., Two Putative c-Type Multiheme Cytochromes Required for the Expression of OmcB, an Outer Membrane Protein Essential for Optimal Fe(III) Reduction in Geobacter sulfurreducens, J. Bacteriol., 2006, vol. 188, pp. 3138–3142.

    Article  PubMed  CAS  Google Scholar 

  8. Lloyd, J.R., Leang, C., Hodges-Myerson, A.L., Coppi, M.V., Cuifo, S., Methe, B., Sandler, S.J., and Lovley, D.R., Biochemical and Genetic Characterization of PpcA, a Periplasmic c-Type Cytochrome in Geobacter sulfurreducens, Biochem. J., 2003, vol. 369, pp. 153–161.

    Article  PubMed  CAS  Google Scholar 

  9. Lovley, D.R, Holmes, D.E, and Nevin, K.P, Dissimilatory Fe(III) and Mn(IV) Reduction, in Advances in Microbial Physiology, Poole, R.K., Ed., London: Elsevier Academic, 2004, pp. 219–286.

    Google Scholar 

  10. Mehta, T., Coppi, M.V., Childers, S.E., and Lovley, D.R., Outer Membrane c-Type Cytochromes Required for Fe(III) and Mn(IV) Oxide Reduction in Geobacter sulfurreducens, Appl. Environ. Microbiol., 2005, vol. 71, pp. 8634–8641.

    Article  PubMed  CAS  Google Scholar 

  11. Pokkuluri, P.R., Londer, Y.Y., Duke, N.E.C., Long, W.C., and Schiffer, M., Family of Cytochrome c 7-Type Proteins from Geobacter sulfurreducens: Structure of One Cytochrome c 7 at 1.45 Å Resolution, Biochemistry, 2004, vol. 43, pp. 849–859.

    Article  PubMed  CAS  Google Scholar 

  12. Pokkuluri, P.R., Londer, Y.Y., Duke, N.E.C., Erickson, J., Pessanha, M., Salgueiro, C.A., and Schiffer, M., Structure of a Novel c 7-Type Three-Heme Cytochrome Domain from a Multidomain Cytochrome c Polymer, Protein Sci. originally published online May, vol. 13, pp. 1684–1692.

  13. Heitmann, D. and Einsle, O., Structural and Biochemical Characterization of DHC2, a Novel Diheme Cytochrome c from Geobacter sulfurreducens, Biochemistry, 2005, vol. 44, pp. 12411–12419.

    Article  PubMed  CAS  Google Scholar 

  14. Pessanha, M., Londer, Y.Y., Long, W.C., Ericson, J., Pokkuluri, P.R., Schiffer, M., and Salgueiro, C.A., Redox Characterization of Geobacter sulfurreducens Cytochrome c 7: Physiological Relevance of the Conserved Residue F15 Probed by Site-Specific Mutagenesis, Biochemistry, 2004, vol. 43, pp. 9909–9917.

    Article  PubMed  CAS  Google Scholar 

  15. Pessanha, M., Morgado, L., Louro, R.O., Londer, Y.Y., Pokkuluri, P.R., Schiffer, M., and Salgueiro, C.A., Thermodinamic Characterization of Triheme Cytochrome PpcA from Geobacter sulfurreducens: Evidence for a Role Played Ione-/H+ Energy Transduction, Biochemistry, 2006, vol. 45, pp. 13910–13917.

    Article  PubMed  CAS  Google Scholar 

  16. Galouchko, A.S., Obraztsova, A.Ya., Shtarkman, N.B., Laurinavichyus, K.S., and Akimenko, V.K., Anaerobic Acetate-Oxidizing Bacterium Transforming Methacrylic Acid, DAN. Seriya Biologicheskaya, 1994, vol. 335, no. 3, pp. 382–384.

    Google Scholar 

  17. Mikulinskaya (Arkhipova), O.V., Galouchko A.S., Laurinavichyus K.S., and Akimenko V.K. Investigation of Methacrylate Reduction by a New Anaerobic Bacterium, Sb. trudov gor. nauch. konf. molodykh uchenykh (Proc. Town Young Scientists Conf.), Pushchino: ONTI PNTs RAN, 1996.

    Google Scholar 

  18. Arkhipova, O.V. and Akimenko, V.K., Unsaturated Organic Acids as Terminal Electron Acceptors for Reductase Chains of Anaerobic Bacteria, Mikrobiologiya, 2005, vol. 74, no. 6, pp. 725–737 [Microbiology (Engl. Transl.), vol. 74, no. 6, pp. 629–639].

    CAS  Google Scholar 

  19. Galouchko, A.S., Mikulinskaya, O.V., Laurinavichyus, K.S., Obraztsova A.Ya., Akimenko V.K., Periplasmic Location of Methacrylate Reductase in Cells of the Strictly Anaerobic Bacterial Strain AM-1, Mikrobiologiya, 1996, vol. 65, no. 4, pp. 495–498 [Microbiology (Engl. Transl.), vol. 65, no. 4, pp. 432–435].

    Google Scholar 

  20. Mikoulinskaia (Arkhipova), O.V., Akimenko, V.K., Galushko, A.S., Thauer, R.K., and Hedderich, R., Cytochrome c-dependent methacrylate reductase from Geobacter sulfurreducens AM-1, Eur. J. Biochem., 1999, vol. 263, pp. 346–352.

    Article  PubMed  CAS  Google Scholar 

  21. Yamanaka, T., C Group Cytochromes, in Metalloproteins. Chemical Properties and Biological Effects. Bioactive Molecules, Otsuka, S. and Yamanaka, T., Eds., Tokyo: Kodanska LTD, 1988, pp. 139–153.

    Google Scholar 

  22. Van der Westen, H.M., Mayhew, S.G., and Veeger, C., Separation of Hydrogenase from Intact Cells of Desulfovibrio vulgaris, FEBS Lett., 1978, vol. 86, no. 1, pp. 122–126.

    Article  PubMed  Google Scholar 

  23. Bradford, M.M., A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem., 1976, vol. 72, pp. 248–254.

    Article  PubMed  CAS  Google Scholar 

  24. Laemmli, U.K., Cleavage of Structural Proteins During the Assembly of the Head of Bacteriophage T4, Nature, 1970, vol. 227, pp. 680–685.

    Article  PubMed  CAS  Google Scholar 

  25. Thomas, P.E., Ruan, D., and Levin, W., An Improved Staining Procedure for the Detection of the Peroxidase Activity of Cytochrome P-450 in Sodium Dodecyl Sulfate Polyacrylamide Gels, Anal. Biochem., 1976, vol. 75, no. 1, pp. 168–176.

    Article  PubMed  CAS  Google Scholar 

  26. Beacham, I.R., Periplasmic Enzymes in Gram-Negative Bacteria, Int. J. Biochem., 1979, vol. 10, no. 11, pp. 877–883.

    Article  PubMed  CAS  Google Scholar 

  27. Watt, S.R. and Clarke, A.J., Role of Autolysins in the EDTA-Induced Lysis of Pseudomonas aeruginosa, FEMS Microbiol. Letts., 1994, vol. 124, pp. 113–120.

    Article  CAS  Google Scholar 

  28. Pilishina, A.V. and Arkhipova, O.V., Isolation and Purification of a Unique Methacrylate Reductase from Geobacter sulfurreducens AM-1, in Sb. tez. 10-i mezhdunar. Pushchinskoi shkoly-konf. molodykh uchenykh “Biologiya — nauka XXI veka” (Proc. 10th Int. Pushchino Conf. of Young Scientists “Biology: Science of the XXI Century”), Pushchino: ONTI PNTs RAN, 2006.

    Google Scholar 

  29. Gross, R., Simon, J., and Kroger, A., Periplasmatic Methacrylate Reductase Activity in Wolinella succinogenes, Arch. Microbiol., 2001, vol. 176, pp. 310–313.

    Article  PubMed  CAS  Google Scholar 

  30. Simon, J., Gross, R., Klimmek, O., Ringel, M., and Kroeger, A., A Periplasmic Flavoprotein in Wolinella succinogenes That Resembles the Fumarate Reductase of Shewanella putrefaciens, Arch. Microbiol., 1998, vol. 169, pp. 424–433.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, Pushchino, Moscow oblast, 142290, Russia

    O. V. Arkhipova, M. S. Chuvochina & S. M. Trutko

Authors
  1. O. V. Arkhipova
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. S. Chuvochina
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. M. Trutko
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O. V. Arkhipova.

Additional information

Original Russian Text © O.V. Arkhipova, M.S. Chuvochina, S.M. Trutko, 2009, published in Mikrobiologiya, 2009, Vol. 78, No. 3, pp. 334–341.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Arkhipova, O.V., Chuvochina, M.S. & Trutko, S.M. Cytochromes c of the anaerobic methacrylate reducer Geobacter sulfurreducens AM-1. Microbiology 78, 296–303 (2009). https://doi.org/10.1134/S0026261709030060

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026261709030060

Key words

Search

Navigation