title: Structure-activity relationships of wheat flavone O-methyltransferase: a homodimer of convenience.
abstract: Wheat flavone O-methyltransferase catalyzes three sequential methylations of the flavone tricetin. Like other flavonoid O-methyltransferases, the protein is a homodimer. We demonstrate, using analytical ultracentrifugation, that perchlorate dissociates the dimer into monomers. The resulting monomers retain all their catalytic capacity, including the ability to catalyze the three successive methylations. We show, using isothermal titration calorimetry, that the binding constant for S-adenosyl-L-methionine does not change significantly as the protein dissociates. The second substrate, tricetin, binds to the dimers but could not be tested with the monomers. CD, UV and fluorescence spectroscopy show that there are substantial changes in the structure of the protein as it dissociates. The fact that there are differences between the monomers and dimers even as the monomers maintain activity may be the result of the very low catalytic capacity of this enzyme. Maximal turnover numbers for the dimers and monomers are only about 6-7 per minute. Even though the binding pockets for S-adenosyl-L-methionine, tricetin, selgin and tricin are intact, selection of a catalytically competent structure may be a very slow step during catalysis.
authors.affiliation: Enzyme Research Group, Department of Biology, Concordia University, Montreal, QC, Canada. krnbltt@alcor.concordia.ca
authors.forename: Jack A
authors.initials: JA
authors.lastname: Kornblatt
authors.forename: Jian-Min
authors.initials: JM
authors.lastname: Zhou
authors.forename: Ragai K
authors.initials: RK
authors.lastname: Ibrahim
mesh_terms: D002867:Chromones / Q000737:chemistry / Q000378:metabolism; D019281:Dimerization; D047309:Flavones / Q000378:metabolism*; D008745:Methylation; D008780:Methyltransferases / Q000737:chemistry*; D008958:Models, Molecular; D015394:Molecular Structure; D008970:Molecular Weight; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014908:Triticum / Q000201:enzymology*
chemical_list: D002867:Chromones; D047309:Flavones; C469689:tricetin; D008780:Methyltransferases