5-demethylubiquinone-9-methyltransferase from rat liver mitochondria. Characterization, localization, and solubilization.
The methyltransferase responsible for the conversion of 5-demethylubiquinone-9 to ubiquinone-9 in rat liver mitochondria has been shown to be localized in the inner membrane of rat liver mitochondria. NADH was required to generate the hydroquinone, which was the immediate substrate for methylation. The Km for 5-demethylubiquinone-9 was estimated to be in the range of 60 to 80 nM and the Km for S-adenosylmethionine was found to be 22 micronM. The methyl-transferase was solubilized by Triton X-100, a procedure which inactivated the 5-demethylubiquinone-9 reductase. Dithionite was found to partially substitute for NADH in both membranous and soluble systems. Inhibitors of catechol-O-methyltransferase were not effective inhibitors of 5-demethylubiquinone-9-methyltransferase. In addition, catechol-O-methyltransferase and 5-demethylubiquinone-9-methyltransferase were found to have reciprocal subcellular localizations. It is likely that the hydrophobic side chain of ubiquinone, added to p-hydroxybenzoate in the  first biosynthetic step, is required for attachment to the lipid bilayer. This permits subsequent metabolism of the ring system by membrane-bound enzymes, including the final methylation to form ubiquinone-9.
D000818:Animals; D002394:Catechol O-Methyltransferase / Q000032:analysis; D002396:Catechols / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D008780:Methyltransferases / Q000302:isolation & purification*; D008930:Mitochondria, Liver / Q000201:enzymology*; D051381:Rats; D012995:Solubility; D014451:Ubiquinone / Q000031:analogs & derivatives
D002396:Catechols; D003902:Detergents; D014451:Ubiquinone; C020004:5-demethylubiquinone-9-methyltransferase; D008780:Methyltransferases; D002394:Catechol O-Methyltransferase