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Crystal structure of VnfH, the iron protein component of vanadium nitrogenase

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Abstract

Nitrogenases catalyze the biological fixation of inert N2 into bioavailable ammonium. They are bipartite systems consisting of the catalytic dinitrogenase and a complementary reductase, the Fe protein that is also the site where ATP is hydrolyzed to drive the reaction forward. Three different subclasses of dinitrogenases are known, employing either molybdenum, vanadium or only iron at their active site cofactor. Although in all these classes the mode and mechanism of interaction with Fe protein is conserved, each one encodes its own orthologue of the reductase in the corresponding gene cluster. Here we present the 2.2 Å resolution structure of VnfH from Azotobacter vinelandii, the Fe protein of the alternative, vanadium-dependent nitrogenase system, in its ADP-bound state. VnfH adopts the same conformation that was observed for NifH, the Fe protein of molybdenum nitrogenase, in complex with ADP, representing a state of the functional cycle that is ready for reduction and subsequent nucleotide exchange. The overall similarity of NifH and VnfH confirms the experimentally determined cross-reactivity of both ATP-hydrolyzing reductases.

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Abbreviations

EPR:

Electron paramagnetic resonance

MR:

Molecular replacement

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Acknowledgements

The authors thank the staff at beam line X06DA of the Swiss Light Source, Paul Scherrer Institute, Villigen, CH, for their excellent assistance with diffraction data collection. This work was supported by the Deutsche Forschungsgemeinschaft (RTG 1976 and PP 1927) and the European Research Council (Grant no. 310656).

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Authors and Affiliations

  1. Institute for Biochemistry, Albert-Ludwigs-University Freiburg, Albertstrasse 21, 79104, Freiburg, Germany

    Michael Rohde, Christian Trncik, Daniel Sippel, Stefan Gerhardt & Oliver Einsle

  2. BIOSS Centre for Biological Signalling Studies, Schänzlestr. 1, 79104, Freiburg, Germany

    Oliver Einsle

Authors
  1. Michael Rohde
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  2. Christian Trncik
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  3. Daniel Sippel
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  4. Stefan Gerhardt
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  5. Oliver Einsle
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Correspondence to Oliver Einsle.

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Rohde, M., Trncik, C., Sippel, D. et al. Crystal structure of VnfH, the iron protein component of vanadium nitrogenase. J Biol Inorg Chem 23, 1049–1056 (2018). https://doi.org/10.1007/s00775-018-1602-4

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  • DOI: https://doi.org/10.1007/s00775-018-1602-4

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