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Electron Paramagnetic Resonance Spectroscopy

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Nitrogen Fixation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 766))

Abstract

EPR spectroscopy has been an important tool in nitrogenase research for the last 50 years. The three metalloclusters in nitrogenase, the Fe protein [4Fe–4S] cluster, and the MoFe protein P-cluster, and FeMo-cofactor, all have EPR spectra when poised in the appropriate paramagnetic states. EPR spectroscopy can probe changes in the electronic properties of each metal cluster, such as when substrates bind, and can provide a definitive method for observing changes in the redox states of the clusters. In this chapter, the methods for analysis of the three metal clusters of nitrogenase by EPR spectroscopy are described, along with methods for trapping substrate-derived intermediates on the active site that are amenable to characterization by EPR and other magnetic resonance spectroscopy techniques.

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Acknowledgments

The authors acknowledge the long collaboration with the Brian Hoffman and Dennis Dean laboratories in advancing understanding of nitrogenase. Work in the laboratory of the authors is supported by a generous grant from the National Institutes of Health (GM59087). The authors also acknowledge the help and guidance from Dr. Brian Bennett of the National Biomedical EPR Center for EPR simulations.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Utah State University, Logan, UT, 84322, USA

    Karamatullah Danyal, Zhi-Yong Yang & Lance C. Seefeldt

Authors
  1. Karamatullah Danyal
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  2. Zhi-Yong Yang
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  3. Lance C. Seefeldt
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Corresponding author

Correspondence to Lance C. Seefeldt .

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Editors and Affiliations

  1. , Dept Molecular Biology & Biochemistry, University of California, Irvine, Irvine, 92697, California, USA

    Markus W. Ribbe

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Danyal, K., Yang, ZY., Seefeldt, L.C. (2011). Electron Paramagnetic Resonance Spectroscopy. In: Ribbe, M. (eds) Nitrogen Fixation. Methods in Molecular Biology, vol 766. Humana Press. https://doi.org/10.1007/978-1-61779-194-9_13

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  • DOI: https://doi.org/10.1007/978-1-61779-194-9_13

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-193-2

  • Online ISBN: 978-1-61779-194-9

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