title: Novel characteristics of UDP-glucose dehydrogenase activities in maize: non-involvement of alcohol dehydrogenases in cell wall polysaccharide biosynthesis.
abstract: UDP-glucose dehydrogenase (UDPGDH) activity was detected in extracts of maize cell-cultures and developing leaves. The reaction product was confirmed as UDP-glucuronate. Leaf extracts from null mutants defective in one or both of the ethanol dehydrogenase genes, ADH1 and ADH2, had similar UDPGDH activities to wild-type, showing that UDPGDH activity is not primarily due to ADH proteins. The mutants showed no defect in their wall matrix pentose:galactose ratios, or matrix:cellulose ratio, showing that ADHs were not required for normal wall biosynthesis. The majority of maize leaf UDPGDH activity had K (m) (for UDP-glucose) 0.5-1.0 mM; there was also a minor activity with an unusually high K (m) of >50 mM. In extracts of cultured cells, kinetic data indicated at least three UDPGDHs, with K (m) values (for UDP-glucose) of roughly 0.027, 2.8 and >50 mM (designated enzymes E(L), E(M) and E(H) respectively). E(M) was the single major contributor to extractable UDPGDH activity when assayed at 0.6-9.0 mM UDP-Glc. Most studies, in other plant species, had reported only E(L)-like isoforms. Ethanol (100 mM) partially inhibited UDPGDH activity assayed at low, but not high, UDP-glucose concentrations, supporting the conclusion that at least E(H) activity is not due to ADH. At 30 microM UDP-glucose, 20-150 microM UDP-xylose inhibited UDPGDH activity, whereas 5-15 microM UDP-xylose promoted it. In conclusion, several very different UDPGDH isoenzymes contribute to UDP-glucuronate and hence wall matrix biosynthesis in maize, but ADHs are not responsible for these activities.
authors.affiliation: The Edinburgh Cell Wall Group, Institute of Molecular Plant Sciences, School of Biological Sciences, The University of Edinburgh, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Kärkönen
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Fry
mesh_terms: D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D007700:Kinetics; D009154:Mutation; D018515:Plant Leaves / Q000201:enzymology; D011134:Polysaccharides / Q000096:biosynthesis*; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000378:metabolism*; D003313:Zea mays / Q000201:enzymology*
chemical_list: D011134:Polysaccharides; D000426:Alcohol Dehydrogenase; D014533:Uridine Diphosphate Glucose Dehydrogenase