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Activity and Structural Changes of Euphorbia characias Peroxidase in the Presence of Trifluoroethanol

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Abstract

Activity assays, conformational changes and transitional switches between secondary structures of a peroxidase from Euphorbia characias were studied in the presence of trifluoroethanol and in the presence or absence of calcium ions. The addition of trifluoroethanol up to 10–20% first induced a drastic decrease of α-helix content followed by an increase of tryptophan fluorescence emission intensity, a progressive re-induction of the formation of α-helical elements concomitant with loss of enzyme activity. In the presence of calcium ions, the fluorescence of the enzyme almost remained unchanged in the trifluoroethanol concentration range 5–20%. Further increase in trifluoroethanol concentration led to a protein structure characterized by a progressive re-induction of α-helical elements, a remarkable increase of the tryptophan fluorescence and a loss of enzyme activity. These results indicate that calcium ions in Euphorbia peroxidase play an essential role in maintaining the hydrophobic interactions on the protein structure preserving enzymatic activity.

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Abbreviations

ABTS:

2,2′-Azinobis(3-ethylbenzthiazoline-6-sulfonic acid)

ELP:

Euphorbia latex peroxidase

TFE:

2,2,2-Trifluoroethanol

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Acknowledgments

This study was partially supported by PRIN 2006 (Progetti di ricerca di interesse nazionale) funds and by a grant from “Fondazione Banco di Sardegna”, Sassari.

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Authors and Affiliations

  1. Department of Applied Sciences in Biosystems, University of Cagliari Cittadella Universitaria, 09042, Monserrato, CA, Italy

    F. Pintus, A. Mura, A. Contini, D. Spanò, R. Medda & G. Floris

  2. Department of Biomedical Sciences and Technologies, University of Cagliari Cittadella Universitaria, 09042, Monserrato, CA, Italy

    A. C. Rinaldi

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  1. F. Pintus
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  2. A. Mura
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  3. A. C. Rinaldi
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  4. A. Contini
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  5. D. Spanò
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  6. R. Medda
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  7. G. Floris
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Correspondence to G. Floris.

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Pintus, F., Mura, A., Rinaldi, A.C. et al. Activity and Structural Changes of Euphorbia characias Peroxidase in the Presence of Trifluoroethanol. Protein J 27, 434–439 (2008). https://doi.org/10.1007/s10930-008-9153-0

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  • DOI: https://doi.org/10.1007/s10930-008-9153-0

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