Characterization of bispecific caffeic acid/5-hydroxyferulic acid O-methyltransferase from aspen.
Bispecific O-methyltransferase (OMT, EC 2.1.1.68) which catalyses the meta-specific methylation of caffeic acid and 5-hydroxyferulic acid was purified to homogeneity from the developing secondary xylem of aspen (Populus tremuloides). The enzyme was purified by conventional techniques and affinity chromatography on S-adenosyl-L-homocysteine agarose using substrate elution. The enzyme has a M(r) of 40,000 as determined by SDS-PAGE. Amino acid sequences of three peptides produced from a proteolytic digest of bispecific OMT were obtained by automated Edman degradation. Polyclonal antibodies raised against the purified OMT reacted strongly to OMT in both purified and unpurified fractions in western blots. Addition of an equal concentration of anti-OMT IgG to crude extract protein inhibited OMT activity by more than 70%.
Phytotechnology Research Center, School of Forestry and Wood Products, Michigan Technological University, Houghton 49931.
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Bugos
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Chiang
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Campbell
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002109:Caffeic Acids / Q000378:metabolism*; D003373:Coumaric Acids / Q000378:metabolism*; D008780:Methyltransferases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology; D013379:Substrate Specificity
D002109:Caffeic Acids; D003373:Coumaric Acids; C071744:5-hydroxyferulic acid; D008780:Methyltransferases; C019608:caffeate O-methyltransferase; C040048:caffeic acid