Methylation of sulfhydryl groups: a new function for a family of small molecule plant O-methyltransferases

Authors

  • Heather Coiner,

    1. TU München, FG Biomolekulare Lebensmitteltechnologie, Lise-Meitner-Str. 34, D-85354 Freising, Germany,
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  • Gudrun Schröder,

    1. Universität Freiburg, Institut für Biologie II, Schänzlestr. 1, D-79104 Freiburg, Germany,
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  • Elke Wehinger,

    1. Universität Freiburg, Institut für Biologie II, Schänzlestr. 1, D-79104 Freiburg, Germany,
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  • Chang-Jun Liu,

    1. Biology Department, Bldg. 463, Brookhaven National Laboratory, 50 Bell Avenue, Upton, NY 11973, USA, and
    2. Howard Hughes Medical Institute, The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
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  • Joseph P. Noel,

    1. Howard Hughes Medical Institute, The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA
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  • Wilfried Schwab,

    1. TU München, FG Biomolekulare Lebensmitteltechnologie, Lise-Meitner-Str. 34, D-85354 Freising, Germany,
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  • Joachim Schröder

    Corresponding author
    1. Universität Freiburg, Institut für Biologie II, Schänzlestr. 1, D-79104 Freiburg, Germany,
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*(fax +49 761 203 2601; e-mail joachim.schroeder@biologie.uni-freiburg.de).

Summary

In plants, type I and II S-adenosyl-l-methionine-dependent O-methyltransferases (OMTs) catalyze most hydroxyl group methylations of small molecules. A homology-based RT-PCR strategy using Catharanthus roseus (Madagascar periwinkle) RNA previously identified six new type I plant OMT family members. We now describe the molecular and biochemical characterization of a seventh protein. It shares 56–58% identity with caffeic acid OMTs (COMTs), but it failed to methylate COMT substrates, and had no activity with flavonoids. However, the in vitro incubations revealed unusually high background levels without added substrates. A search for the responsible component revealed that the enzyme methylated dithiothreitol (DTT), the reducing agent added for enzyme stabilization. Unexpectedly, product analysis revealed that the methylation occurred on a sulfhydryl moiety, not on a hydroxyl group. Analysis of 34 compounds indicated a broad substrate range, with a preference for small hydrophobic molecules. Benzene thiol (Km 220 μm) and furfuryl thiol (Km 60 μm) were the best substrates (6–7-fold better than DTT). Small isosteric hydrophobic substrates with hydroxyl groups, like phenol and guaiacol, were also methylated, but the activities were at least 5-fold lower than with thiols. The enzyme was named C. roseus S-methyltransferase 1 (CrSMT1). Models based on the COMT crystal structure suggest that S-methylation is mechanistically identical to O-methylation. CrSMT1 so far is the only recognized example of an S-methyltransferase in this protein family. Its properties indicate that a few changes in key residues are sufficient to convert an OMT into a S-methyltransferase (SMT). Future functional investigations of plant methyltransferases should consider the possibility that the enzymes may direct methylation at sulfhydryl groups.

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