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X-ray structural studies of the fungal laccase from Cerrena maxima

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Abstract

Laccases are members of the blue multi-copper oxidase family. These enzymes oxidize substrate molecules by accepting electrons at a mononuclear copper centre and transferring them to a trinuclear centre. Dioxygen binds to the trinuclear centre and following the transfer of four electrons is reduced to two molecules of water. The X-ray structure of a laccase from Cerrena maxima has been elucidated at 1.9 Å resolution using synchrotron data and the molecular replacement technique. The final refinement coefficients are Rcryst = 16.8% and Rfree = 23.0%, with root mean square deviations on bond lengths and bond angles of 0.015 Å and 1.51°, respectively. The type 1 copper centre has an isoleucine residue at the axial position and the “resting” state of the trinuclear centre comprises a single oxygen (OH) moiety asymmetrically disposed between the two type 3 copper ions and a water molecule attached to the type 2 ion. Several carbohydrate binding sites have been identified and the glycan chains appear to promote the formation of well-ordered crystals. Two tyrosine residues near the protein surface have been found in a nitrated state.

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Notes

  1. In crystallographic terms an occupancy of 50% means that in the large number of molecules that constitute the crystal used for the data collection only 50% possess a copper ion at the respective site; the remaining molecules have unoccupied sites. The occupancy does not determine the oxidation state of the metal ion.

  2. Recently some 50% of the C. maxima sequence has become available (V. Tishkov, Moscow University, Russia). These incomplete data show a 98% identity with the sequence derived from the electron density maps.

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Acknowledgements

This project was supported by BMBF (Bundesministerium fur Forshung and Wissenschaft under contract no. RUS/214). We would like to thank the Komarov Botanic Institute of the Russian Academy of Sciences (St. Peterburg) for generously donating the C. maxima fungus strain. We also thank the EMBL Outstation, Hamburg, for access to the synchrotron radiation facilities at DESY where the X-ray data collection was undertaken. Figures 1, 2, 3 and 4 were produced using the program PyMOL [55].

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Authors and Affiliations

  1. A.V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninskiy Prospect 59, 119333, Moscow, Russia

    Andrey V. Lyashenko, Nadezhda E. Zhukhlistova, Yuliya N. Zhukova, Azat G. Gabdoulkhakov, Ekaterina Y. Morgunova & Al`bert M. Mikhailov

  2. Instituto de Tecnologia Química e Biológica (ITQB), Universidade Nova de Lisboa, Apartado 127, Av. Republica, 2781-901, Oeiras, Portugal

    Isabel Bento & Peter F. Lindley

  3. Centre of Biomolecular Sciences, University of St. Andrews, North Hough, St. Andrews Fife, KY16 9ST, UK

    Viatcheslav N. Zaitsev

  4. Institute of Biochemistry, Tuebingen University, Hoppe-Seyler-Strasse 4, 72076, Tübingen, Germany

    Wolfgang Voelter

  5. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Institutskaya Street 3, 142290, Puschino, Moscow Region, Russia

    Galina S. Kachalova

  6. A.N. Bakh Institute of Biochemistry, Russian Academy of Sciences, Leninskiy Prospect 33, 119071, Moscow, Russia

    Elena V. Stepanova & Ol`ga V. Koroleva

  7. European Molecular Biology Laboratory, c/o DESY, Notkestrasse 85, 22603, Hamburg, Germany

    Victor S. Lamzin

  8. Department of Chemical Enzymology, M.V. Lomonosov Moscow State University, 119992, Moscow, Russia

    Vladimir I. Tishkov

  9. Institute fur Biochemie und Lebensmittelchemie, University of Hamburg, c/o DESY, Building 22, Notkestrasse 85, 22603, Hamburg, Germany

    Christian Betzel

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  1. Andrey V. Lyashenko
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  2. Isabel Bento
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  3. Viatcheslav N. Zaitsev
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  14. Christian Betzel
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  15. Peter F. Lindley
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  16. Al`bert M. Mikhailov
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Corresponding authors

Correspondence to Viatcheslav N. Zaitsev, Peter F. Lindley or Al`bert M. Mikhailov.

Additional information

The diffraction data and the model coordinates have been deposited into the Protein Data Bank with accession code ID PDB 2H5U.

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Lyashenko, A.V., Bento, I., Zaitsev, V.N. et al. X-ray structural studies of the fungal laccase from Cerrena maxima . J Biol Inorg Chem 11, 963–973 (2006). https://doi.org/10.1007/s00775-006-0158-x

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  • DOI: https://doi.org/10.1007/s00775-006-0158-x

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