Carbone tetrachloride, silymarin, as well as other chemicals, Colla-genase, 1-chloro-2,4-dinitrobenzene
elicited increase in vacuolar glutathione-S-conjugate transport activity.
Reduced glutathione (GSH), 1-chloro-2,4-dinitrobenzene
(CDNB), reduced nicotinamide adenine dinucleotide phosphate (NADPH), amidopyrine, cytochrome C, thiobarbituric acid and all other chemicals were purchased from Sigma Chemical Company (Saint Louis, USA).
GST activity was assayed using 1-chloro-2,4-dinitrobenzene
reagent (CDNB) (Habig et al.
Among spectrophotometric determinations, the GSH5,5'-dithio-bis(2-nitrobenzoic acid) "recycling assay" (40) and the glutathione S-transferase 1-chloro-2,4-dinitrobenzene
(CDNB) endpoint method (41) are the most commonly used.
The active sites of the two structures, which were quite different, represented snapshots along the reaction coordinate for the enzyme-catalyzed reaction of glutathione with 1-chloro-2,4-dinitrobenzene
and revealed specific interactions between the enzyme, intermediate, and product that are important for catalysis.
The activity of GST was determined using 1-chloro-2,4-dinitrobenzene
as a substrate (Habig et al.
The glutathione S-transferase activity was measured using the 1-chloro-2,4-dinitrobenzene
We quantified the resorufin formation using a resorufin standard curve and measured the activity of GST by monitoring GST-dependent conjugation of glutathione and 1-chloro-2,4-dinitrobenzene
spectrophotometrically using a model UV-2401 PC spectrophotometer (Shimadzu, Columbia, MD) at 340 nm (Habig et al.
The reagents used for the experiments included [alpha]-naphthylacetate ([alpha]-NA), Eserine, AlbuMin bovine V (BSA), Coomassie Brilliant Blue G250, 1-chloro-2,4-dinitrobenzene
(CDNB),reduced glutathione (GSH), Fast Blue B salt, Sodium dodecyl sulfate (SDS), EDTA, 1,4-dithioerythritol (DTT), phenylmethanesulfonyl fluoride (PMSF) and 4-nitroanisole (PNA),which were all bought from Sigma.
The principle of the GST activity assay is based upon the GST-catalysed reaction between GSH and GST substrate, 1-chloro-2,4-dinitrobenzene
(CDNB) as described by Habig et al.
GST activity was determined on the basis of conjugation of GSH with 1-chloro-2,4-dinitrobenzene
(CDNB) (Habig et al.