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A concerted mechanism for berberine bridge enzyme

Nature Chemical Biology volume 4, pages 739–741 (2008)Cite this article

Abstract

Berberine bridge enzyme catalyzes the conversion of (S)-reticuline to (S)-scoulerine by formation of a carbon-carbon bond between the N-methyl group and the phenolic ring. We elucidated the structure of berberine bridge enzyme from Eschscholzia californica and determined the kinetic rates for three active site protein variants. Here we propose a catalytic mechanism combining base-catalyzed proton abstraction with concerted carbon-carbon coupling accompanied by hydride transfer from the N-methyl group to the N5 atom of the FAD cofactor.

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Figure 1: Reaction and structure of BBE.

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References

  1. Steffens, P., Nagakura, N. & Zenk, M.H. Phytochem. 24, 2577–2583 (1985).

    Article  CAS  Google Scholar 

  2. Kutchan, T.M. & Dittrich, H. J. Biol. Chem. 270, 24475–24481 (1995).

    Article  CAS  Google Scholar 

  3. Edmondson, D.E., Binda, C. & Mattevi, A. Arch. Biochem. Biophys. 464, 269–276 (2007).

    Article  CAS  Google Scholar 

  4. Scrutton, N.S. Nat. Prod. Rep. 21, 722–730 (2004).

    Article  CAS  Google Scholar 

  5. Fitzpatrick, P.F. Bioorg. Chem. 32, 125–139 (2004).

    Article  CAS  Google Scholar 

  6. Winkler, A., Hartner, F., Kutchan, T.M., Glieder, A. & Macheroux, P. J. Biol. Chem. 281, 21276–21285 (2006).

    Article  CAS  Google Scholar 

  7. Huang, C.-H. et al. J. Biol. Chem. 280, 38831–38838 (2005).

    Article  CAS  Google Scholar 

  8. Alexeev, I., Sultana, A., Mantsala, P., Niemi, J. & Schneider, G. Proc. Natl. Acad. Sci. USA 104, 6170–6175 (2007).

    Article  CAS  Google Scholar 

  9. Heuts, D.P.H.M., Winter, R.T., Damsma, G.E., Janssen, D.B. & Fraaije, M.W. Biochem. J. 413, 175–183 (2008).

    Article  CAS  Google Scholar 

  10. Rand, T., Qvist, K.B., Walter, C.P. & Poulsen, C.H. FEBS J. 273, 2693–2703 (2006).

    Article  CAS  Google Scholar 

  11. Harayama, T., Tezuka, Y., Taga, T. & Yoneda, F. J. Chem. Soc. [Perkin 1] 75–83 (1987).

  12. Iwata, M., Bruice, T.C., Carrell, H.L. & Glusker, J.P. J. Am. Chem. Soc. 102, 5036–5044 (1980).

    Article  CAS  Google Scholar 

  13. Settembre, E.C. et al. Biochemistry 42, 2971–2981 (2003).

    Article  CAS  Google Scholar 

  14. Malito, E., Coda, A., Bilyeu, K.D., Fraaije, M.W. & Mattevi, A. J. Mol. Biol. 341, 1237–1249 (2004).

    Article  CAS  Google Scholar 

  15. Miller, J.R. & Edmondson, D.E. Biochemistry 38, 13670–13683 (1999).

    Article  CAS  Google Scholar 

  16. Bjorklund, J.A. et al. J. Am. Chem. Soc. 117, 1533–1545 (1995).

    Article  CAS  Google Scholar 

  17. Winkler, A., Kutchan, T.M. & Macheroux, P. J. Biol. Chem. 282, 24437–24443 (2007).

    Article  CAS  Google Scholar 

  18. Sirikantaramas, S. et al. J. Biol. Chem. 279, 39767–39774 (2004).

    Article  CAS  Google Scholar 

  19. Taura, F. et al. FEBS Lett. 581, 2929–2934 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We appreciate the support of staff scientists at the synchrotron beamlines at DESY/EMBL-Hamburg during diffraction data collection and of Hansjörg Weber (Institute of Organic Chemistry, Graz University of Technology) for recording the NMR spectra. Financial support was provided by the Austrian Science Fund (Fonds zur Förderung der wissenschaftlichen Forschung) through the Doktoratskolleg “Molecular enzymology” W901-B05 (to K.G. and P.M.).

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Author notes

  1. Andrzej Łyskowski & Sabrina Riedl

    Present address: Present addresses: Institute of Biotechnology, PO Box 65, Viikinkaari 1, FIN-00014 University of Helsinki, Finland (A.L.) and Institute of Biophysics and Nanosystems Research, Austrian Academy of Sciences, Schmiedlstraße 6, 8042 Graz, Austria (S.R.).,

Authors and Affiliations

  1. Institute of Biochemistry, Graz University of Technology, Petersgasse 12/II, Graz, 8010, Austria

    Andreas Winkler, Sabrina Riedl, Martin Puhl & Peter Macheroux

  2. Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/III, Graz, 8010, Austria

    Andrzej Łyskowski & Karl Gruber

  3. Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, 63132, Missouri, USA

    Toni M Kutchan

Authors
  1. Andreas Winkler
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  2. Andrzej Łyskowski
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  3. Sabrina Riedl
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  5. Toni M Kutchan
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Contributions

A.W., P.M. and K.G. designed experiments and analyzed results. A.W., P.M. and K.G. wrote the manuscript. A.W., A.L. and K.G. determined the crystal structures. A.W., S.R. and M.P. generated, expressed and purified mutant proteins. A.W. and M.P. performed enzyme kinetic experiments. T.M.K. prepared and provided special compounds and helped in editing the manuscript.

Corresponding authors

Correspondence to Sabrina Riedl or Karl Gruber.

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Supplementary Text and Figures

Supplementary Figures 1–7, Supplementary Table 1, Supplementary Discussion and Supplementary Methods (PDF 749 kb)

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Winkler, A., Łyskowski, A., Riedl, S. et al. A concerted mechanism for berberine bridge enzyme. Nat Chem Biol 4, 739–741 (2008). https://doi.org/10.1038/nchembio.123

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