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Chemical Synthesis of an Asymmetric Mimic of the Nitrogenase Active Site

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Metalloproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1876))

Abstract

The synthetic inorganic chemistry of metal–sulfur (M-S, M = metals) clusters has played an important, complementary role to the biochemical analyses of nitrogenase toward a better understanding of the enzyme active site. The active site of nitrogenase (designated the M-cluster) can be extracted from the protein in a solvent-stabilized form, [(cit)MoFe7S9C] (cit = (R)-homocitrate). One important finding of the extracted M-cluster is its catalytic activity toward the reduction of C1-substrates (CN−, CO, CO2) into C1–C5 hydrocarbons in solution. This catalytic property poses challenges for chemists to reproduce the function with synthetic mimics, not only because of the biochemical interests but also due to the potential significance in green chemistry and catalysis research. In this context, our successful synthesis of an asymmetric Mo-Fe-S cluster, [Cp*MoFe5S9(SH)]3−, is one of the recent important achievements in synthetic M-S chemistry, as this cluster catalyzes the reduction of C1-substrates in a similar manner to the extracted M-cluster. Even though the synthetic protocol for this cluster has been described in the literature, there are plenty of pitfalls for researchers unfamiliar with synthetic M-S chemistry. In this chapter, we provide general precautionary statements and detailed protocols for the synthesis of [Cp*MoFe5S9(SH)]3−, with a brief discussion of the experimental tips based on the authors’ experience in both biochemical and synthetic chemical fields.

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Acknowledgments

This work was financially supported by Takeda Science Foundation, the Hori Sciences and Arts Foundation, and Grant-in-Aids for Scientific Research (No. 16H04116) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (to Y.O.). The authors thank Dr. Nathaniel S. Sickerman and Lee Rettberg (University of California, Irvine) for proofreading and fruitful discussions.

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA, USA

    Kazuki Tanifuji

  2. Department of Chemistry, Graduate School of Science, Nagoya University, Nagoya, Japan

    Yasuhiro Ohki

Authors
  1. Kazuki Tanifuji
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  2. Yasuhiro Ohki
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Corresponding authors

Correspondence to Kazuki Tanifuji or Yasuhiro Ohki .

Editor information

Editors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of California-Irvine, Irvine, CA, USA

    Yilin Hu

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Tanifuji, K., Ohki, Y. (2019). Chemical Synthesis of an Asymmetric Mimic of the Nitrogenase Active Site. In: Hu, Y. (eds) Metalloproteins. Methods in Molecular Biology, vol 1876. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8864-8_15

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  • DOI: https://doi.org/10.1007/978-1-4939-8864-8_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8863-1

  • Online ISBN: 978-1-4939-8864-8

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