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Chemical Synthesis of an Asymmetric Mimic of the Nitrogenase Active Site

  • Kazuki Tanifuji
  • Yasuhiro Ohki
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Synthetic mimics Iron–molybdenum–sulfur clusters Nitrogenase Asymmetry Air-free techniques 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Molecular Biology and BiochemistryUniversity of California, IrvineIrvineUSA
  2. 2.Department of Chemistry, Graduate School of ScienceNagoya UniversityNagoyaJapan

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