Membrane-associated reactions in ubiquinone biosynthesis: 2-octaprenyl-3-methyl-5-hydroxy-6-methoxy-1,4-benzoquinone methyltransferase

• Published in: Biochimica et biophysica acta Vol. 428; no. 1; pp. 146 - 156
• Main Authors: Leppik, Ray A., Stroobant, Paul, Shineberg, Barry, Young, Ian G., Gibson, Frank
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 25.03.1976
• Subjects: Animals; Cations, Divalent; Cell Membrane - enzymology; Cell Membrane - metabolism; Centrifugation, Density Gradient; Chromatography, Gel; Dithiothreitol - metabolism; Escherichia coli - enzymology; Genes; Methyltransferases - isolation & purification; Methyltransferases - metabolism; Molecular Weight; Mutation; Quinones; Transduction, Genetic; Ubiquinone - biosynthesis; OMHMB
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/0304-4165(76)90116-1

The O-methylation of 2-octaprenyl-3-methyl-5-hydroxy-6-methoxy-1,4-benzoquinone, which has been previously postulated to be the final reaction in the biosynthesis of ubiquinone was demonstrated in vitro using cell extracts of Escherichia coli. S- Adenosyl- l- methionine was active as the methyl donor for the reaction. The enzyme concerned, S-adenosyl- l-methionine: 2-octaprenyl-3-methyl-5-hydroxy-6-methoxy-1,4-benzoquinone- O- methyltransferase, was partially purified and shown to have a molecular weight of about 50 000 and to require a divalent metal and dithiothreitol for optimal acitivity in vitro. The methyltransferase was absent from extracts from ubiG − mutants suggesting that the ubiG gene is the structural gene coding for the methyltransferase. The enzyme, although not firmly membrane-bound, showed some affinity for the cell membrane in broken cell preparations and could utilize the benzoquinone substrate when the latter was free or bound to the cell membrane, with about equal efficiency. It is concluded that in vivo, the methyltransferase reaction probably occurs at the internal surface of the cytoplasmic membrane.