title: Control of activity through oxidative modification at the conserved residue Cys66 of aryl sulfotransferase IV.
abstract: Oxidation at Cys66 of rat liver aryl suflotransferase IV alters the enzyme's catalytic activity, pH optima and substrate specificity. Although this is a cytosolic detoxification enzyme, the pH optimum for the standard assay substrate 4-nitrophenol is at pH 5.5; upon oxidation, the optimum changes to the physiological pH range. The principal effect of the change in pH optimum is activation, which is manifest by an increase in K'cat without any major influence on substrate binding. In contrast, with tyrosine methyl ester as a substrate, the enzyme's optimum activity occurs at pH 8.0; upon oxidation, it ceases to be a substrate at any pH. The presence of Cys66 was essential for activation to occur, thereby providing a putative reason underlying the conserved nature of this cysteine throughout the phenol sulfotransferase family. Mapping of disulfides by mass spectrometry showed the critical event to be the oxidation of Cys66 to form a disulfide with either Cys232 or glutathione, either one is effective. These results point to a mechanism for regulating the activity of a key enzyme in xenobiotic detoxication during cellular oxidative stress.
authors.affiliation: Section on Enzymes, Laboratory of Biochemistry and Metabolism, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: A D
authors.initials: AD
authors.lastname: Marshall
authors.forename: J F
authors.initials: JF
authors.lastname: Darbyshire
authors.forename: A P
authors.initials: AP
authors.lastname: Hunter
authors.forename: P
authors.initials: P
authors.lastname: McPhie
authors.forename: W B
authors.initials: WB
authors.lastname: Jakoby
mesh_terms: D000818:Animals; D003545:Cysteine; D004220:Disulfides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005978:Glutathione / Q000031:analogs & derivatives / Q000494:pharmacology; D019803:Glutathione Disulfide; D000441:Hexanols / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology; D013058:Mass Spectrometry; D009596:Nitrophenols / Q000494:pharmacology; D010084:Oxidation-Reduction; D010085:Oxidative Phosphorylation; D010449:Peptide Mapping; D051381:Rats; D015238:Sulfotransferases / Q000737:chemistry*
chemical_list: D004220:Disulfides; D000441:Hexanols; D009596:Nitrophenols; C036260:1-hexanol; D015238:Sulfotransferases; C077557:tyrosine-ester sulfotransferase; D005978:Glutathione; D003545:Cysteine; D019803:Glutathione Disulfide; C024836:4-nitrophenol