Comparison of the effects of bile acids and GSH on the fluorescence of bound 1-anilino-8-naphthalene sulfonate and the enzymatic activity of cationic and neutral human hepatic GSH S-transferases.
Three cationic (C1, C2, A1) and a neutral (N1) glutathione (GSH) S-transferase were purified to homogeneity from human liver, as we have previously reported. GSH had no effect on the fluorescence of 1-anilino-8-naphthalene sulfonate (ANS) bound by transferase C1 and N1, but markedly enhanced the fluorescence with C2 and A1 without changing the affinity for ANS. This effect of GSH was saturable and with C2 was intermediate between A1 and C1. Bile acids inhibited the fluorescence of ANS bound to C1 and C2. GSH in the presence of bile acids further decreased the fluorescence of ANS bound to C1 and increased the fluorescence with C2. Transferase A1 showed decreased fluorescence in the presence of lithocholic acid and increased fluorescence in the presence of cholic acid; both changes were reversed by GSH. Transferase N1 showed increased fluorescence of bound ANS in the presence of various bile acids and this effect was diminished in the presence of GSH. Enzyme activity of the transferase was inhibited by bile acids with the exception of transferase A1. All the proteins bound lithocholic acid. The inhibition of C1 and N1 was greater at pH 6.5 than 7.4 and the order of addition of substrates and inhibitor made no difference.
Takikawa
H
H
Liver Research Laboratory, Wadsworth Veterans Administration Hospital Center, Los Angeles.
Sugiyama
Y
Y
Kaplowitz
N
N
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism*; D001647:Bile Acids and Salts / Q000378:metabolism / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology*; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology
D000817:Anilino Naphthalenesulfonates; D001647:Bile Acids and Salts; D005456:Fluorescent Dyes; C027132:1-anilino-8-naphthalenesulfonate; D005982:Glutathione Transferase; D005978:Glutathione