Spectrofluorimetric assays for rat liver phenol sulphotransferases I/II and III/IV based on a common substrate

A spectrofluorimetric method for the separate assay of two subgroups of rat liver phenol sulphotransferases (PSTs I and II and PSTs III and IV) has been established. The method is based on the enzyme reaction using two different concentrations of p-hydroxybenzaldehyde (10 μM for PSTs III/IV and 3 mM for PSTs I/II) as the substrate and the spectrofluorimetric determination of the product p-formylphenyl sulphate by means of its reaction with 1,4-dimethyl-3-carbamoylpyridinium chloride, a selective reagent for aromatic aldehydes without free phenolic hydroxy groups. The detection limit of the sulphate is 50–70 pmol in the enzyme reaction mixutre. The K_m values for p-hydroxybenzaldehyde with PSTs I/II and III/IV were 1.1 mM and 2.1 μM, respectively. The effects of growth and of dexamethasone treatment of rats on the liver PST activities were also studied.