Purification, crystallization and preliminary X-ray study of the fungal laccase from Cerrena maxima
- A. V. Shubnikov Institute of Crystallography, RAS, Leninskiy Prospect 59, 119333 Moscow (Russian Federation)
- Institute of Biochemistry, University of Tuebingen, Physiologisch-Chemisches Institut, Hoppe-Seyler-Strasse 4, 72076 Tuebingen (Germany)
- University of St Andrews, Centre for Biomolecular Sciences, North Haugh, St Andrews, KY16 9ST,Scotland (United Kingdom)
- Instituto de Tecnologia Química e Biológica (ITQB), Universidade Nova de Lisboa, Apartado 127, Av. Republica, 2781-901 Oeiras (Portugal)
- A. N. Bakh Institute of Biochemistry, RAS, Leninskiy Prospect 33, 119071 Moscow (Russian Federation)
- Institute of Theoretical and Experimental Biophysics of RAS, Institutskaya Street 3, 142290 Puschino, Moscow Region (Russian Federation)
- Department of Chemical Enzymology, M. V. Lomonosov Moscow State University, 119992 Moscow (Russian Federation)
- European Molecular Biology Laboratory, c/o DESY, Notkestrasse 85, 22603 Hamburg (Germany)
- University of Hamburg, Institute fur Biochemie und Lebensmittelchemie, Department of Biochemistry and Molecular Biology, c/o DESY, Building 22a, Notkestrasse 85, 22603 Hamburg (Germany)
The crystallization and preliminary X-ray structure at 1.9 Å resolution of the fungal laccase from C. maxima are presented. Laccases are members of the blue multi-copper oxidase family that 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. Crystals of the laccase from Cerrena maxima have been obtained and X-ray data were collected to 1.9 Å resolution using synchrotron radiation. A preliminary analysis shows that the enzyme has the typical laccase structure and several carbohydrate sites have been identified. The carbohydrate chains appear to be involved in stabilization of the intermolecular contacts in the crystal structure, thus promoting the formation of well ordered crystals of the enzyme. Here, the results of an X-ray crystallographic study on the laccase from the fungus Cerrena maxima are reported. Crystals that diffract well to a resolution of at least 1.9 Å (R factor = 18.953%; R{sub free} = 23.835; r.m.s.d. bond lengths, 0.06 Å; r.m.s.d. bond angles, 1.07°) have been obtained despite the presence of glycan moieties. The overall spatial organization of C. maxima laccase and the structure of its copper-containing active centre have been determined by the molecular-replacement method using the laccase from Trametes versicolor (Piontek et al., 2002 ▶) as a structural template. In addition, four glycan-binding sites were identified and the 1.9 Å X-ray data were used to determine the previously unknown primary structure of this protein. The identity (calculated from sequence alignment) between the C. maxima laccase and the T. versicolor laccase is about 87%. Tyr196 and Tyr372 show significant extra density at the ortho positions and this has been interpreted in terms of NO{sub 2} substituents.
- OSTI ID:
- 22356374
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 10; Other Information: PMCID: PMC2225196; PMID: 17012782; PUBLISHER-ID: pu5149; OAI: oai:pubmedcentral.nih.gov:2225196; Copyright (c) International Union of Crystallography 2006; This is an open-access article distributed under the terms described at http://journals.iucr.org/services/termsofuse.html.; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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