Skip to main content
SpringerLink
Log in

Probing the stability of the disulfide radical intermediate of thioredoxin using direct electrochemistry

  • Published:
Letters in Peptide Science Aims and scope Submit manuscript

Abstract

Thioredoxin, a redox active disulfide protein, has been specifically immobilized at a modified gold electrode. The thioredoxin is uniquely oriented relative to the electrode surface via a histidine tag thereby enabling the redox mechanism of protein to be examined. When scanning the applied potential in the negative direction (cathodic), two one-electron reduction waves can be observed. The first of these redox waves occurs at −90 mV and is electrochemically reversible at all scan rates whereas the second wave occurs at −433 mV is irreversible. These two processes are interpreted as the initial reduction of the disulfide form of the protein to a stable (reversible) semi-reduced radical anion intermediate, followed by an electrochemically irreversible process to form a fully reduced thioredoxin. These electron transfer characteristics suggest that a radical intermediate retaining the sulfur-sulfur bond is thermodynamically stable but the addition of a second electron results in bond scission.

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. Arner, E.S. J. and Holmgren, A., Eur. J. Biochem., 267 (2000) 6102.

    Article  CAS  PubMed  Google Scholar 

  2. Watson, W.H., Pohl, J., Montfort, W.R., Stuchlik, O., Reed, M.S., Powis, G. and Jones, D.P., J. Biol. Chem., 278 (2003) 33408.

    CAS  PubMed  Google Scholar 

  3. Sevier, C. S. and Kaiser, C.A., Nat. Rev., 3 (2002) 836.

    Article  CAS  Google Scholar 

  4. Das, K. C. and White, C.W., Proc. Nat. Acad. Sci., 99 (2002) 9617.

    CAS  PubMed  Google Scholar 

  5. El Hanine Lmoumene, C., Conte, D. and Houee-Levin, C., Biochem., 39 (2000) 9295.

    CAS  Google Scholar 

  6. Houee-Levin, C., Meth. Enzym., 353 (2002) 35.

    CAS  PubMed  Google Scholar 

  7. Chivers, P.T., Prehoda, K. E. and Raines, R.T., Biochemistry, 36 (1997) 4061.

    CAS  PubMed  Google Scholar 

  8. Wynn, R., Cocco, M. J. and Richards, F.M., Biochemistry, 34 (1995) 11807.

    Article  CAS  PubMed  Google Scholar 

  9. Hirasawa, M., Schurmann, P., Jacquot, J-P., Manieri, W., Jacquot, P., Keryer, E., Hartman, F. C. and Knaff, D.B., Biochemistry, 38 (1999) 5200.

    Article  CAS  PubMed  Google Scholar 

  10. Setterdahl, A.T., Chivers, P.T., Hirasawa, M., Lemaire, S.D., Keryer, E., Miginiac-Maslow, M., Kim, S.-K., Mason, J., Jacquot, J.-P., Longbine, C.C., de Lamotte-Guery, F. and Knaff, D.B., Biochemistry, 42 (2003) 14877.

    Article  CAS  PubMed  Google Scholar 

  11. Mossner, E., Huber-Wunderlich, M., Rietsch, A., Beckwith, J., Glockshuber, R. and Aslund, F., J. Biol. Chem., 274 (1999) 25254.

    Article  CAS  PubMed  Google Scholar 

  12. Krause, G., Lundstrom, J., Barea, J.L., de la Cuesta P. and Holmgren, A., J Biol. Chem., 266 (1991) 9494.

    CAS  PubMed  Google Scholar 

  13. Salamon, Z., Tollin, G., Hirasawa, M., Gardet-Salvi, L., Stritt-Etter, A.-L., Knaff, D. B. and Schurmann, P., Biochim. Biophys. Acta114 (1995) 1230.

    Google Scholar 

  14. Salamon, Z., Gleason, F. K. and Tollin, G., Arch. Biochem. Biophys., 299 (1992) 193.

    Article  CAS  PubMed  Google Scholar 

  15. Martin, L.L., Johnson, D. L. and Rodgers, R., Aust. Prov. Patent Number 2003903504, (2003).

  16. Johnson, D. L. and Martin, L.L., (2004), manuscript in preparation.

  17. Johnson, D. L. and Martin, L.L., Langmuir, (2004) manuscript in preparation.

  18. Johnson, D.L., Thompson, J.L., Brinkmann, S., Schuller, K. A. and Martin, L.L., Biochemistry, 10 (2003) 10229.

    Google Scholar 

  19. Johnson, D.L., Norman, S., Tuckey, R. C. and Martin, L.L. Bioelectrochemistry, 59 (2003) 41.

    Article  CAS  PubMed  Google Scholar 

  20. Johnson, D.L., Maxwell, C.J. Losic, D., Shapter, J.G. and Martin, L.L., Bioelectrochemistry, 58 (2002) 137.

    Article  CAS  PubMed  Google Scholar 

  21. Laviron, E., J. Electroanal. Chem., 101 (1979) 19.

    Article  CAS  Google Scholar 

  22. Gauduel, Y., Gelabert, H. and Guilloud, F., J. Am. Chem. Soc., 122 (2000) 5082.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Chemistry, Physics and Earth Sciences, Flinders University, South Australia, 5001, Australia

    Daniel L. Johnson & Lisandra L. Martin

  2. The University of Adelaide, South Australia, 5005, Australia

    Steven W. Polyak & John C. Wallace

Authors
  1. Daniel L. Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Steven W. Polyak
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John C. Wallace
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lisandra L. Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Johnson, D.L., Polyak, S.W., Wallace, J.C. et al. Probing the stability of the disulfide radical intermediate of thioredoxin using direct electrochemistry. Int J Pept Res Ther 10, 495–500 (2003). https://doi.org/10.1007/s10989-004-2410-y

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10989-004-2410-y

Search

Navigation