Activation and detoxication of N-hydroxy-Trp-P-2 by glutathione and glutathione transferases

Abstract

The roles of non-enzymatic and enzymatic glutathione (GSH) conjugation in the activation and detoxication of 3-hydroxy-amino-1-methyl-5H-pyrido[4,3-b]indole (N-OH-Trp-P-2) were studied in vitro. N-OH-Trp-P-2 is an active metabolite of 3-amino-1-methyl-5H-pyrido[4,3-d]indole (Trp-P-2), a mutagenic and carcinogenic heterocyclic amine. 3-Nitroso-1-methyl-5H-pyrido[4,3-b]indole (NO-Trp-P-2) reacted rapidly and non-enzymatically with GSH to form N-OH-Trp-P-2 and a small amount of two GSH conjugates (CN-1 and CN-2). On the other hand, non-enzymatic reaction of GSH with N-OH-Trp-P-2 was very slow, but the GSH conjugation with N-OH-Trp-P-2 was catalyzed by rat liver GSH transferase and a rat liver cytosol fraction to form three conjugates (CH-1, CH-2 and CH-3). The enzymatic conjugation was effectively inhibited by organic tin compounds which are known as powerful GSH transferase inhibitors. The conjugates were unstable enough to yield Trp-P-2 (from CN-1, CN-2 and CH-2) or N-OH-Trp-P-2 (from CH-3) on incubation at 37 degrees C for 30-60 min. Only CH-1 was stable under similar conditions. The mutagenicities of the GSH conjugates and the effects of GSH and GSH transferase were studied by using Salmonella typhimurium TA98 as the tester strain. The GSH conjugates except for CH-3 were completely detoxicated products, but CH-3 was found to be a more potent mutagen than N-OH-Trp-P-2. The mutagenicity of CH-3 seemed to be due to the direct action of the conjugate, and not to N-OH-Trp-P-2 formed from it.