Inducible UDP-glucose dehydrogenase from French bean (Phaseolus vulgaris L.) locates to vascular tissue and has alcohol dehydrogenase activity.
UDP-glucose dehydrogenase is responsible for channelling UDP-glucose into the pool of UDP-sugars utilized in the synthesis of wall matrix polysaccharides and glycoproteins. It has been purified to homogeneity from suspension-cultured cells of French bean by a combination of hydrophobic-interaction chromatography, gel filtration and dye-ligand chromatography. The enzyme had a subunit of Mr 40,000. Km values were measured for UDP-glucose as 5.5 +/- 1.4 mM and for NAD+ as 20 +/- 3 microM. It was subject to inhibition by UDP-xylose. UDP-glucose dehydrogenase activity co-purified with alcohol dehydrogenase activity from suspension-cultured cells, elicitor-treated cells and elongating hypocotyls, even when many additional chromatographic steps were employed subsequently. The protein from each source was resolved into virtually identical patterns of isoforms on two-dimensional isoelectric focusing/PAGE. However, a combination of peptide mapping and sequence analysis, gel analysis using activity staining and kinetic analysis suggests that both activities are a function of the same protein. An antibody was raised and used to immunolocalize UDP-glucose dehydrogenase to developing xylem and phloem of French bean hypocotyl. Together with data published previously, these results are consistent with an important role in the regulation of carbon flux into wall matrix polysaccharides.
Robertson
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Department of Biochemistry, Royal Holloway and Bedford New College, University of London, Egham, Surrey, U.K.
Smith
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C
Bolwell
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GP
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism*; D000906:Antibodies; D004790:Enzyme Induction; D007887:Fabaceae / Q000201:enzymology*; D007150:Immunohistochemistry; D007700:Kinetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000302:isolation & purification* / Q000378:metabolism*; D010946:Plants, Medicinal; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000096:biosynthesis* / Q000302:isolation & purification*
D000906:Antibodies; D010940:Plant Proteins; D000426:Alcohol Dehydrogenase; D014533:Uridine Diphosphate Glucose Dehydrogenase