Brief Communication | Published:

A concerted mechanism for berberine bridge enzyme

Nature Chemical Biology volume 4, pages 739741 (2008) | Download Citation

Subjects

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.

  • Compound

    (S)-Reticuline

  • Compound

    Berberine

  • Compound

    Sanguinarine

  • Compound

    (S)-Scoulerine

  • Compound

    Flavin adenine dinucleotide

  • Compound

    (2R,3S,4S)-5-[(4R)-6',7'-Dimethyl-2,3',5-trioxo-1'H-spiro[imidazolidine-4,2'-quinoxalin]-4'(3'H)-yl]-2,3,4-trihydroxypentyl-adenosine-diphosphate

  • Compound

    (R,S)-Laudanosine

  • Compound

    6,7-Dimethoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline

  • Compound

    Formaldehyde

  • Compound

    (S)-N-Methylcoclaurine

  • Compound

    (S)-Coclaurine

  • Compound

    (S)-Coreximine

  • Compound

    2-Methyl-3,4-dihydropapaverinium

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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.).

Author information

Author notes

    • Andrzej Łyskowski
    •  & Sabrina Riedl

    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.).

Affiliations

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

    • Andreas Winkler
    • , Sabrina Riedl
    • , Martin Puhl
    •  & Peter Macheroux
  2. Institute of Molecular Biosciences, University of Graz, Humboldtstraße 50/III, 8010 Graz, Austria.

    • Andrzej Łyskowski
    •  & Karl Gruber
  3. Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, Missouri 63132, USA

    • 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.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–7, Supplementary Table 1, Supplementary Discussion and Supplementary Methods

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DOI

https://doi.org/10.1038/nchembio.123

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