Assay of purified aryl sulfotransferase suitable for reactions yielding unstable sulfuric acid esters.
An assay procedure for purified aryl sulfotransferase is described. The method utilizes isocratic paired-ion reverse-phase HPLC analysis of adenosine-3',5'-diphosphate formed in the reaction. Evaluation of the assay procedure was carried out with 1-naphthalene-methanol as a model substrate for purified rat hepatic aryl sulfotransferase IV. Kinetic constants for sulfation of 1-naphthalenemethanol determined by this method compared favorably with those determined using thin-layer chromatographic assays of 35S incorporation. These results indicate that the method will be suitable for determination of kinetic constants in sulfotransferase-catalyzed reactions where the product sulfuric acid ester may be chemically unstable.
Duffel
M W
MW
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Binder
T P
TP
Rao
S I
SI
D000244:Adenosine Diphosphate / Q000032:analysis; D000818:Animals; D015239:Arylsulfotransferase / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D005069:Evaluation Studies as Topic; D006868:Hydrolysis; D008297:Male; D009281:Naphthalenes / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D009281:Naphthalenes; C050116:1-naphthalenemethanol; D000244:Adenosine Diphosphate; D015239:Arylsulfotransferase