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N-terminal domains mediate [2Fe-2S] cluster transfer from glutaredoxin-3 to anamorsin

Nature Chemical Biology volume 11, pages 772–778 (2015)Cite this article

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Abstract

In eukaryotes, cytosolic monothiol glutaredoxins are proteins implicated in intracellular iron trafficking and sensing via their bound [2Fe-2S] clusters. We define a new role of human cytosolic monothiol glutaredoxin-3 (GRX3) in transferring its [2Fe-2S] clusters to human anamorsin, a physical and functional protein partner of GRX3 in the cytosol, whose [2Fe-2S] cluster–bound form is involved in the biogenesis of cytosolic and nuclear Fe-S proteins. Specific protein recognition between the N-terminal domains of the two proteins is the mandatory requisite to promote the [2Fe-2S] cluster transfer from GRX3 to anamorsin.

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Figure 1: Cluster transfer from GRX3 to anamorsin.
Figure 2: Impaired cluster transfer and no protein-protein interaction between GRX3(GrxA/B) and anamorsin.
Figure 3: Apo-GRX3 and apo-anamorsin specifically recognize each other via their N-terminal domains.
Figure 4: N domains of apo-GRX3 and of apo-anamorsin form a heterodimer stabilized through the unstructured linker of anamorsin.
Figure 5: Working model for the functional role of cluster transfer from GRX3 to anamorsin in iron metabolism.

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Acknowledgements

We thank A. Gallo (CERM) for assistance in recording the EPR spectra. This work was supported by Ente Cassa di Risparmio (Grant ID no. 2013/7101); the Ministero dell'Istruzione, dell'Università e della Ricerca (Grant ID number: CTN01_00177_962865); the European Integrated Structural Biology Infrastructure (INSTRUCT), which is part of the European Strategy Forum on Research Infrastructures; and national member subscriptions.

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Authors and Affiliations

  1. Magnetic Resonance Center CERM, University of Florence, Florence, Italy

    Lucia Banci, Simone Ciofi-Baffoni, Karolina Gajda, Riccardo Muzzioli, Riccardo Peruzzini & Julia Winkelmann

  2. Department of Chemistry, University of Florence, Florence, Italy

    Lucia Banci, Simone Ciofi-Baffoni & Riccardo Muzzioli

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  1. Lucia Banci
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Contributions

The work plan was conceived and designed by L.B. and S.C.-B.; R.M. and K.G. produced protein constructs of anamorsin and GRX3; L.B. and S.C.-B. planned the experiments; J.W. and R.P. performed and analyzed NMR data; K.G., J.W. and R.M. performed and analyzed EPR and UV-vis experiments. The manuscript was drafted by L.B. and S.C.-B. and revised by all authors.

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Correspondence to Lucia Banci or Simone Ciofi-Baffoni.

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Supplementary Results, Supplementary Tables 1 and 2 and Supplementary Figures 1–8. (PDF 5454 kb)

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Banci, L., Ciofi-Baffoni, S., Gajda, K. et al. N-terminal domains mediate [2Fe-2S] cluster transfer from glutaredoxin-3 to anamorsin. Nat Chem Biol 11, 772–778 (2015). https://doi.org/10.1038/nchembio.1892

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