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An artificial di-iron oxo-protein with phenol oxidase activity

Nature Chemical Biology volume 5, pages 882–884 (2009)Cite this article

Abstract

Here we report the de novo design and NMR structure of a four-helical bundle di-iron protein with phenol oxidase activity. The introduction of the cofactor-binding and phenol-binding sites required the incorporation of residues that were detrimental to the free energy of folding of the protein. Sufficient stability was, however, obtained by optimizing the sequence of a loop distant from the active site.

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Figure 1: Comparison of DF1 and DF3 structures.
Figure 2: Model of substrate interaction with DF3 protein.

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Acknowledgements

The authors thank M. Trifuoggi for metal content analysis. This work was supported by the Italian Ministry of University and Scientific Research (PRIN 2007KAWXCL), the European Commission (Marie Curie Fellowship HPMD-GH-01-00113-03 to R.T.M.R.), the US National Institutes of Health (GM54616) and the US National Science Foundation (grant from the Materials Research Science and Engineering Center to the Laboratory for Research on the Structure of Matter at the University of Pennsylvania).

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  1. Rafael Torres Martin de Rosales & Ornella Maglio

    Present address: Present addresses: Division of Imaging Sciences, King's College London, St. Thomas Hospital, London, UK (R.T.M.R.) and Institute of Biostructure and Bioimaging, National Research Council (CNR), Napoli, Italy (O.M.).,

Authors and Affiliations

  1. Department of Chemistry, University Federico II of Napoli, Complesso Universitario Monte S. Angelo, Napoli, Italy

    Marina Faiella, Concetta Andreozzi, Rafael Torres Martin de Rosales, Vincenzo Pavone, Ornella Maglio, Flavia Nastri & Angela Lombardi

  2. Department of Biochemistry and Biophysics, University of Pennsylvania, School of Medicine, Philadelphia, Pennsylvania, USA

    William F DeGrado

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  1. Marina Faiella
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  2. Concetta Andreozzi
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  3. Rafael Torres Martin de Rosales
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  4. Vincenzo Pavone
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Contributions

F.N., O.M. and A.L. designed experiments and analyzed results. R.T.M.R., M.F. and C.A. synthesized the protein and performed UV-vis and chemical denaturation studies. O.M. and M.F. determined the solution structure. F.N. and C.A. performed kinetic experiments. A.L and V.P. performed molecular modeling. W.F.D. and A.L. wrote the manuscript in consultation with the remaining authors.

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Correspondence to William F DeGrado or Angela Lombardi.

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Faiella, M., Andreozzi, C., de Rosales, R. et al. An artificial di-iron oxo-protein with phenol oxidase activity. Nat Chem Biol 5, 882–884 (2009). https://doi.org/10.1038/nchembio.257

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