title: A spectrophotometric glutathione S-transferase assay displaying alpha-class selectivity utilizing 1-p-chlorophenyl-4,4-dimethyl-5-diethylamino-1-penten-3- one hydrobromide.
abstract: The conjugation of 1-p-chlorophenyl-4,4-dimethyl-5-diethylamino-1-penten-3- one hydrobromide (CDDP), a Mannich base of an alpha,beta-unsaturated ketone, to glutathione is catalyzed selectively by alpha-class glutathione S-transferase. The reaction of CDDP with glutathione can be monitored continuously by following the conjugation-dependent decrease in absorbance of CDDP at 307 nm. The Km and Vmax for CDDP with alpha-class glutathione S-transferase from horse liver were determined to be 226 microM and 14.6 mumol/(min.mg), respectively. CDDP is the first example of an alpha-class glutathione S-transferase selective substrate that monitors the glutathione conjugation activity, rather than the glutathione peroxidase activity of the enzyme.
authors.affiliation: Department of Chemistry and Biochemistry, University of Windsor, Ont., Canada.
authors.forename: D J
authors.initials: DJ
authors.lastname: Sexton
authors.forename: J R
authors.initials: JR
authors.lastname: Dimmock
authors.forename: B
authors.initials: B
authors.lastname: Mutus
mesh_terms: D002722:Chlorobenzenes / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000032:analysis* / Q000145:classification; D007659:Ketones / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002722:Chlorobenzenes; D007659:Ketones; C059871:1-(4-chlorophenyl)-4,4-dimethyl-5-diethylamino-1-penten-3-one; D005982:Glutathione Transferase; D005978:Glutathione