Arabidopsis glucosyltransferases with activities toward both endogenous and xenobiotic substrates

• Published in: Planta 2003-05, Vol.217 (1), p.138-146
• Main Authors: Meßner, Burkhard, Thulke, Oliver, Schäffner, Anton R.
• Format: Article
• Language: English
• Subjects: Alcohols; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis Proteins - chemistry; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Biochemistry; Biological and medical sciences; Carboxylic acids; Chlorophenols - pharmacology; Coumaric Acids - pharmacology; Detoxification; Enzyme Activation - drug effects; Enzyme Inhibitors - pharmacology; Enzyme substrates; Enzymes; Flavonoids - pharmacology; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic - drug effects; Gene Expression Regulation, Plant - drug effects; Glucosyltransferases - chemistry; Glucosyltransferases - genetics; Glucosyltransferases - metabolism; Hydroxyls; Kaempferols; Kinetics; Metabolism; Metabolism. Physicochemical requirements; Parabens; Phenols; Plant Leaves - enzymology; Plant Leaves - genetics; Plant physiology and development; Plant Roots - enzymology; Plant Roots - genetics; Propionates; Substrate Specificity; Substrates; Xenobiotics; Xenobiotics - metabolism; Xenobiotics - pharmacology
• ISSN: 0032-0935; 1432-2048
• DOI: 10.1007/s00425-002-0969-0

Arabidopsis thaliana Heynh. harbors UDP-glucose-dependent glucosyltransferase (UGT; EC 2.4.1.-) activities that are able to glucosylate xenobiotic substrates as a crucial step in their detoxification, similar to other plants. However, it has remained elusive whether side-activities of UGTs acting on endogenous substrates could account for that property. Therefore, seven recombinantly expressed A. thaliana enzymes were tested using the phytotoxic xenobiotic model compound 2,4,5-trichlorophenol (TCP) as a substrate. The enzymes were selected from the large Arabidopsis UGT gene family because their previously identified putative endogenous substrates comprised both carboxylic acid, and phenolic and aliphatic hydroxyl moieties as biochemical targets. In addition, UGT75D1, which was shown to accept the endogenous flavonoid kaempferol as a substrate, was included. All enzymes tested, except the sterol-conjugating UGT80A2, glucosylated TCP as a parallel activity. The Km values for TCP ranged from 0.059 to 1.25 mM. When tested at saturating concentrations of the native substrates the glucosylation of TCP by the glucose-ester-forming UGT84A1 and UGT84A2 was suppressed by p-coumaric acid and sinapic acid, respectively. In contrast, the activities of UGT72E2 and UGT75D1 toward their phenolic native substrates and the xenobiotic TCP were mutually inhibited. TCP was a competitive inhibitor of sinapyl alcohol glucosylation by UGT72E2. These overlapping in vitro activities suggest cross-talk between the detoxification of xenobiotics and endogenous metabolism at the biochemical level, depending on the presence of competing substrates and enzymes.