Skip to main content
SpringerLink
Log in

ESR Spectroscopy Investigation of the Denaturation Process of Soybean Peroxidase Induced by Guanidine Hydrochloride, DMSO or Heat

  • Published:
The Protein Journal Aims and scope Submit manuscript

Abstract

The involvement of protein denaturation and/or misfolding processes in the insurgence of several diseases raises the interest in structural dynamic studies of proteins. The use of nitroxide spin labels with electron paramagnetic resonance is a powerful tool for detecting structural changes in proteins. In the present study, we apply this strategy to soybean peroxidase (SBP), a protein characterised by high thermal and structural stability, and we propose a simple method to analyse the anisotropy changes of the protein system and to relate them with the structural changes induced by protein unfolding. We examined the effect of temperature, guanidine hydrochloride and dimethylsulfoxide on the stability of SBP and looked for correlations between the ESR results and the experimental findings obtained by other techniques, reported in the literature. The agreement between data obtained through different strategies supports the validity and reliability of the ESR approach to protein unfolding.

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

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.

Similar content being viewed by others

Abbreviations

CD:

circular dichroism

Cm:

melting concentration

DMAB:

3-dimethylamino-benzoic acid

DMSO:

dimethylsulfoxide

ESR:

Electron spin resonance

GuHCl:

guanidine hydrochloride

HRP:

horseradish peroxidase

MBTH:

3-methyl-2-benzothiazolinone hydrazone hydrochloride

SBP:

soybean peroxidase

τC :

correlation time

Tm:

melting temperature

References

  • Cortes D.M., Cuello L.G., Peroso E. (2001). J. Gen. Physiol. 117:165–180

    Article  CAS  Google Scholar 

  • Dunford H.B. (1999). Heme peroxidases. John Wiley & sons, New York

    Google Scholar 

  • Fajer, P. G. (2000). In: Meyers, R. A. (ed.), Encyclopedia of Analytical Chemistry. Wiley & Sons, Chichester, pp. 5725–5761.

  • Henriksen A., Mirza O., Indiani C., Teilum K., Smulevich G., Welinder K., Gajhede R. (2001). Protein Sci. 10:108–15

    Article  CAS  Google Scholar 

  • Hubbell W. L., Cafiso D. S., Altenbach C. (2000). Nat. Struct. Biol. 7:735–739

    Article  CAS  Google Scholar 

  • Hubbell W. L., Mchaourab H. S., Altenbach C., Lietzow M. A. (1996). Structure 4:779–783

    Article  CAS  Google Scholar 

  • Hubbell W. L., Altenbach C. (1994). Curr. Opin. Struc. Biol. 4:566–573

    Article  CAS  Google Scholar 

  • Hurt K. M., Nilges K. M., Carlson K. E., Tamrazi A., Belford K. E., Katzenellenbogen J. A. (2004). Biochemistry 43:1891–1907

    Article  Google Scholar 

  • Hustedt E. J., Beth A. H. (1999). Annu. Rev. Biophys. Biomol. Struct. 28:129–153

    Article  CAS  Google Scholar 

  • Kamal J. K., Nazeerunnisa M., Behere D. V. (2002a). J. Biol. Chem. 277:40717–40721

    Article  CAS  Google Scholar 

  • Kamal J. K., Behere D. V. (2002b). Biochemistry 41:9034–9042

    Article  CAS  Google Scholar 

  • Kamal J. K., Behere D. V. (2003). J. Inorg. Biochem. 94:236–242

    Article  CAS  Google Scholar 

  • McEldoon J. P., Dordick J. S. (1996). Biotechnol. Prog. 12:555–558

    Article  CAS  Google Scholar 

  • Rabenstein M. D., Shin Y. K. (1995). Proc. Natl. Acad. Sci. USA 92:8239–8243

    Article  CAS  Google Scholar 

  • Rose, G. (2002) in: Advances in Protein Chemistry, vol 62, Unfolded proteins, Academic Press.

  • Santucci R., Laurenti E., Sinibaldi F., Ferrari R.P. (2002). Biochim. Biophys. Acta 1596:225–233

    CAS  Google Scholar 

  • Strancar J., Sentjurc M., Schara M. (2000). J. Magn. Res. 142:254–265

    Article  CAS  Google Scholar 

  • Voss J., Hubbel W. L., Kaback H. R. (1995). Proc. Natl. Acad. Sci. USA 92:12300–12303

    Article  CAS  Google Scholar 

  • Welinder K., Larsen Y. B. (2004). Biochim. Biophys. Acta 1698:121–126

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Dip. Chimica I.F.M., Università di Torino, Via Giuria 7, 10125, Torino, Italy

    Barbara Boscolo, Enzo Laurenti & Elena Ghibaudi

Authors
  1. Barbara Boscolo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Enzo Laurenti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elena Ghibaudi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Elena Ghibaudi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Boscolo, B., Laurenti, E. & Ghibaudi, E. ESR Spectroscopy Investigation of the Denaturation Process of Soybean Peroxidase Induced by Guanidine Hydrochloride, DMSO or Heat. Protein J 25, 379–390 (2006). https://doi.org/10.1007/s10930-006-9024-5

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10930-006-9024-5

Keywords

Search

Navigation