Abstract
Copper is essential for proper functioning of cytochrome c oxidases, and therefore for cellular respiration in eukaryotes and many bacteria. Here we show that a new periplasmic protein (PCuAC) selectively inserts Cu(I) ions into subunit II of Thermus thermophilus ba3 oxidase to generate a native CuA site. The purported metallochaperone Sco1 is unable to deliver copper ions; instead, it works as a thiol-disulfide reductase to maintain the correct oxidation state of the CuA cysteine ligands.
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Acknowledgements
This work was supported by the European Commission (European Network of Research Infrastructures for Providing Access and Technological Advancements in Bio-NMR contract n° 026145, SPINE2-Complexes contract n° LSHG-CT-2006-031220 and Marie Curie host fellowships for early stage research training n° MEST-CT-2004-504391, NMR in Inorganic Structural Biology) and by a grant from Ente Cassa di Risparmio di Firenze. Work in Rosario (Argentina) was supported by the US National Institutes of Health (R01-GM068682), the Howard Hughes Medical Institute and Agencia Nacional de Promoción Científica y Tecnológica (PME2003-0026 and PICT2002-01-11625) grants to A.J.V. L.A.A. thanks Consejo Nacional de Investigaciones Científícas y Técnicas for a doctoral fellowship. We thank D. Winge (Departments of Medicine and Biochemistry, University of Utah Health Sciences Center, University of Utah) for kindly providing the expression plasmid for Tt Sco1.
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L.A.A. cloned and expressed Tt Sco1 and Tt CuA proteins and contributed to the NMR structure of Tt Sco1 and NMR titrations; P.G. cloned and expressed PCuAC protein and performed redox experiments; S.W. solved the structures of PCuAC and contributed to the NMR titrations and NMR structure of Tt Sco1; S.C.-B. supervised and coordinated the acquisition of NMR data and structures, and the performance of titration experiments; I.B., L.B., G.A.S. and A.J.V. developed and directed the project and contributed to the writing. All authors were involved in the discussion of the biochemical meaning of the experiments.
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Abriata, L., Banci, L., Bertini, I. et al. Mechanism of CuA assembly. Nat Chem Biol 4, 599–601 (2008). https://doi.org/10.1038/nchembio.110
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DOI: https://doi.org/10.1038/nchembio.110
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