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A [4Fe–4S]-Fe(CO)(CN)-l-cysteine intermediate is the first organometallic precursor in [FeFe] hydrogenase H-cluster bioassembly

Nature Chemistry volume 10, pages 555–560 (2018)Cite this article

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Abstract

Biosynthesis of the [FeFe] hydrogenase active site (the 'H-cluster') requires the interplay of multiple proteins and small molecules. Among them, the radical S-adenosylmethionine enzyme HydG, a tyrosine lyase, has been proposed to generate a complex that contains an Fe(CO)2(CN) moiety that is eventually incorporated into the H-cluster. Here we describe the characterization of an intermediate in the HydG reaction: a [4Fe–4S][(Cys)Fe(CO)(CN)] species, 'Complex A', in which a CO, a CN− and a cysteine (Cys) molecule bind to the unique 'dangler' Fe site of the auxiliary [5Fe–4S] cluster of HydG. The identification of this intermediate—the first organometallic precursor to the H-cluster—validates the previously hypothesized HydG reaction cycle and provides a basis for elucidating the biosynthetic origin of other moieties of the H-cluster.

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Fig. 1: [FeFe] hydrogenase (HydA) H-cluster bioassembly.
Fig. 2: Deconvolution of the HydG reaction EPR spectra.
Fig. 3: 13C Mims-ENDOR spectra of the HydG reaction mixtures generated under the standard condition.
Fig. 4: 57Fe HYSCORE spectra of HydG reaction mixture generated under the standard condition.
Fig. 5: Dicyano analogue of Complex A.

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Acknowledgements

We thank J. Swartz from Stanford University for providing the E. coli strain overexpressing SoHydG. This work is supported by National Institute of Health (GM104543). Correspondence and requests for materials should be addressed to R.D.B. The authors declare no competing financial interests.

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  1. Daniel L. M. Suess

    Present address: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA

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  1. Department of Chemistry, University of California, Davis, CA, USA

    Guodong Rao, Lizhi Tao, Daniel L. M. Suess & R. David Britt

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G.R., L.T., D.L.M.S. and R.D.B. designed the experiments. G.R., L.T. and D.L.M.S. performed the experiments and analysed the data. All the authors contributed to writing the manuscript.

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Correspondence to R. David Britt.

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Rao, G., Tao, L., Suess, D.L.M. et al. A [4Fe–4S]-Fe(CO)(CN)-l-cysteine intermediate is the first organometallic precursor in [FeFe] hydrogenase H-cluster bioassembly. Nature Chem 10, 555–560 (2018). https://doi.org/10.1038/s41557-018-0026-7

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