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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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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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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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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DOI: https://doi.org/10.1038/nchembio.257
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