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An unusual carbon–carbon bond cleavage reaction during phosphinothricin biosynthesis

Nature volume 459, pages 871–874 (2009)Cite this article

Abstract

Natural products containing phosphorus–carbon bonds have found widespread use in medicine and agriculture1. One such compound, phosphinothricin tripeptide, contains the unusual amino acid phosphinothricin attached to two alanine residues. Synthetic phosphinothricin (glufosinate) is a component of two top-selling herbicides (Basta and Liberty), and is widely used with resistant transgenic crops including corn, cotton and canola. Recent genetic and biochemical studies showed that during phosphinothricin tripeptide biosynthesis 2-hydroxyethylphosphonate (HEP) is converted to hydroxymethylphosphonate (HMP)2. Here we report the in vitro reconstitution of this unprecedented C(sp3)–C(sp3) bond cleavage reaction and X-ray crystal structures of the enzyme. The protein is a mononuclear non-haem iron(ii)-dependent dioxygenase that converts HEP to HMP and formate. In contrast to most other members of this family, the oxidative consumption of HEP does not require additional cofactors or the input of exogenous electrons. The current study expands the scope of reactions catalysed by the 2-His–1-carboxylate mononuclear non-haem iron family of enzymes.

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Figure 1: Structure of phosphinothricin tripeptide and the reaction catalysed by HEPD.
Figure 2: NMR and mass spectral data from in vitro labelling studies.
Figure 3: Structures of Cd( ii )–HEPD and Cd( ii )–HEPD–HEP.
Figure 4: Working models for the mechanism of catalysis by HEPD.

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Protein Data Bank

Data deposits

Atomic coordinates and structure factors have been deposited in the Protein Data Bank (PDB) under accession codes 3G7D (for the apo structure) and 3GBF (for the liganded structure).

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Acknowledgements

We thank B. Griffin, J. M. Bollinger, S. E. Denmark and T. Begley for discussions. This work was supported by grants from the National Institutes of Health (PO1 GM077596 to W.W.M., W.A.v.d.D. and S.K.N., and NIH RO1 GM59334 to W.W.M.) and by the University of Illinois.

Authors Contributions R.M.C. performed all biochemical assays shown, which were designed and analysed by R.M.C. and W.A.v.d.D. All structural studies were performed and interpreted by H.Z. and S.K.N. W.W.M. designed and J.A.V.B. performed initial biochemical reactions and identified the products. J.T.W. and G.L. synthesized all substrates. R.M.C., S.K.N. and W.A.v.d.D. wrote the manuscript.

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Author notes

  1. Robert M. Cicchillo and Houjin Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA,

    Robert M. Cicchillo, John T. Whitteck, Gongyong Li & Wilfred A. van der Donk

  2. Institute for Genomic Biology,,

    Robert M. Cicchillo, Houjin Zhang, John T. Whitteck, Satish K. Nair, Wilfred A. van der Donk & William W. Metcalf

  3. Howard Hughes Medical Institute,,

    Robert M. Cicchillo & Wilfred A. van der Donk

  4. Department of Biochemistry, and,

    Houjin Zhang, Satish K. Nair & Wilfred A. van der Donk

  5. Department of Microbiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802, USA,

    Joshua A. V. Blodgett & William W. Metcalf

Authors
  1. Robert M. Cicchillo
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  2. Houjin Zhang
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Corresponding authors

Correspondence to Satish K. Nair, Wilfred A. van der Donk or William W. Metcalf.

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Cicchillo, R., Zhang, H., Blodgett, J. et al. An unusual carbon–carbon bond cleavage reaction during phosphinothricin biosynthesis. Nature 459, 871–874 (2009). https://doi.org/10.1038/nature07972

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