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Tyrosyl Radical in the W164Y Mutant of P. eryngii Versatile Peroxidase: an EPR and DFT/PCM Study

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Abstract

Versatile peroxidases (VP) constitute a new class of high redox potential fungal enzymes that are able to degrade lignin and large substrate molecules. These enzymes catalyze the oxidation of substrates at an exposed tryptophan radical formed by a long-range electron transfer mechanism to heme following the activation by H2O2. In a previous paper, it was demonstrated using electron paramagnetic resonance (EPR) and electron-nuclear double resonance experiments on wild-type VP that Trp164 was the radical site and that it was in a hydrogen-bonded neutral form. In this paper, the W164Y variant is analyzed and it is shown that also the variant is able to form the so-called Compound I (VPI) in the form of protein radical, although in different yields with respect to the wild type. The X-band EPR experiments in combination with density functional theory/polarizable continuum model calculations show that the W164Y mutant is able to form a neutral radical on Tyr164 residue, after activation by H2O2, but in contrast to Trp164, tyrosine is not expected to be hydrogen bonded.

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Acknowledgments

The work was supported by the University of Siena (PAR 2006 to R.P.) and PRIN 234 MIUR (2007 to R.B.).

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Authors and Affiliations

  1. Department of Chemistry, University of Siena, Via A. De Gasperi, 53100, Siena, Italy

    Caterina Bernini, Adalgisa Sinicropi, Riccardo Basosi & Rebecca Pogni

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  1. Caterina Bernini
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  2. Adalgisa Sinicropi
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  3. Riccardo Basosi
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  4. Rebecca Pogni
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Correspondence to Rebecca Pogni.

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Bernini, C., Sinicropi, A., Basosi, R. et al. Tyrosyl Radical in the W164Y Mutant of P. eryngii Versatile Peroxidase: an EPR and DFT/PCM Study. Appl Magn Reson 37, 279–288 (2010). https://doi.org/10.1007/s00723-009-0068-5

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