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Structural snapshots along the reaction pathway of ferredoxin–thioredoxin reductase

Nature volume 448, pages 92–96 (2007)Cite this article

Abstract

Oxygen-evolving photosynthetic organisms regulate carbon metabolism through a light-dependent redox signalling pathway1. Electrons are shuttled from photosystem I by means of ferredoxin (Fdx) to ferredoxin–thioredoxin reductase (FTR), which catalyses the two-electron-reduction of chloroplast thioredoxins (Trxs). These modify target enzyme activities by reduction, regulating carbon flow2. FTR is unique in its use of a [4Fe–4S] cluster and a proximal disulphide bridge in the conversion of a light signal into a thiol signal2. We determined the structures of FTR in both its one- and its two-electron-reduced intermediate states and of four complexes in the pathway, including the ternary Fdx–FTR–Trx complex. Here we show that, in the first complex (Fdx–FTR) of the pathway, the Fdx [2Fe–2S] cluster is positioned suitably for electron transfer to the FTR [4Fe–4S] centre. After the transfer of one electron, an intermediate is formed in which one sulphur atom of the FTR active site is free to attack a disulphide bridge in Trx and the other sulphur atom forms a fifth ligand for an iron atom in the FTR [4Fe–4S] centre—a unique structure in biology. Fdx then delivers a second electron that cleaves the FTR–Trx heterodisulphide bond, which occurs in the Fdx–FTR–Trx complex. In this structure, the redox centres of the three proteins are aligned to maximize the efficiency of electron transfer from the Fdx [2Fe–2S] cluster to the active-site disulphide of Trxs. These results provide a structural framework for understanding the mechanism of disulphide reduction by an iron–sulphur enzyme3 and describe previously unknown interaction networks for both Fdx and Trx (refs 4–6).

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Figure 1: Overall structures of Fdx–FTR, FTR–Trx- f (C49S) and Fdx–FTR–Trx- f (C49S) complexes.
Figure 2: Interactions of Fdx–FTR, FTR–Trx- f (C49S) and Fdx–FTR–Trx- f (C49S) complexes.
Figure 3: FTR mechanism proposed on the basis of current structural and spectroscopic b15 investigations.
Figure 4: Comparison of the active sites of FTR at different reaction states.

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Acknowledgements

We thank J. Kappler, P. Marrack and J. Bolin for support and encouragement; the Zuckerman/Canyon Ranch and A. Lapporte for support of the X-ray and computing facilities; the Howard Hughes Medical Institute beamlines at Advanced Light Source (ALS), the Structural Biology Centre at Advanced Photon Source (APS), and the European Synchrotron Radiation Facility (ESRF) for synchrotron data. H.E. was supported by the Swedish Council for Forestry and Agricultural Research and the Swedish Natural Science Research Council, and P.S. was supported by the Schweizerischer Nationalfonds.

Author Contributions S.D., R.F., D.A.G., F.B., W.M. and P.S. performed the experiments. S.D., R.F., P.S. and H.E. designed and prepared the manuscript.

The atomic coordinates and structure factors of Fdx–FTR, NEM-FTR, two-electron-reduced FTR, FTR–Trx-f(C49S), Fdx–FTR–Trx-f(C49S) and FTR–Trx-m(C40S) have been deposited in the RCSB Protein Data Bank under accession codes 2PVG, 2PUO, 2PVD, 2PU9, 2PVO and 2PUK, respectively.

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Author notes

  1. Rosmarie Friemann, Dominique A. Glauser, Florence Bourquin & Wanda Manieri

    Present address: Present addresses: Department of Chemistry, University of Oslo, PB 1033, Blindern, 0315 Oslo, Norway (R.F.); Fondation pour Recherches Médicales, CH-1211 Genève, Switzerland (D.A.G.); Biochemisches Institut, Universität Zürich, CH-8057 Zürich, Switzerland (F.B.); College of Pharmacy, Medical Center, University of Cincinnati, Cincinnati, Ohio 45267-0004, USA (W.M.).,

Authors and Affiliations

  1. Integrated Department of Immunology, Howard Hughes Medical Institute, National Jewish Medical and Research Center & University of Colorado Health Sciences Center, 1400 Jackson Street, Denver, Colorado 80206, USA,

    Shaodong Dai

  2. Department of Molecular Biology, Swedish University of Agricultural Sciences, Biomedical Centre, Box 590, S-75124 Uppsala, Sweden,

    Rosmarie Friemann & Hans Eklund

  3. Université de Neuchâtel, Laboratoire de Biologie Moléculaire et Cellulaire, Rue Emile Argand 11, CH-2009 Neuchâtel, Switzerland,

    Dominique A. Glauser, Florence Bourquin, Wanda Manieri & Peter Schürmann

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Correspondence to Shaodong Dai.

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Dai, S., Friemann, R., Glauser, D. et al. Structural snapshots along the reaction pathway of ferredoxin–thioredoxin reductase. Nature 448, 92–96 (2007). https://doi.org/10.1038/nature05937

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