Kinetic evidence for the existence of an unstable intermediate in the trinitrophenylation-induced rhodanese inactivation reaction.
1. Rhodanese inactivation by 2,4,6-trinitrobenzenesulphonate, in the presence of n-butylamine in the reaction medium, has been studied by a kinetic analysis of the data, based on the assumption that enzyme inactivation is brought about by direct reaction of this with the modifying agent. 2. Initial reaction rates for rhodanese activity loss were determined by a mathematical analysis of the first three recorded values of rhodanese residual activity. 3. It was found that fractional rhodanese activity values, at infinite reaction time with 2,4,6-trinitrobenzenesulphonate (end-point values), were significantly lower than the values calculated on the assumption of rhodanese inactivation being entirely due to direct trinitrophenylation of enzyme protein. 4. Also, initial enzyme inactivation values were higher in the presence, rather than in the absence, of n-butylamine. 5. These results indicate that 2,4,6-trinitrobenzenesulphonate-induced rhodanese inactivation, in the presence of n-butylamine in the reaction medium, is due to the generation of a highly reactive, unstable intermediate, probably a free radical species.
Rakitzis
E T
ET
Department of Biological Chemistry, University of Athens Medical School, Greece.
Malliopoulou
T B
TB
D000818:Animals; D002082:Butylamines; D002417:Cattle; D004795:Enzyme Stability; D007700:Kinetics; D013884:Thiosulfate Sulfurtransferase / Q000037:antagonists & inhibitors*; D014302:Trinitrobenzenesulfonic Acid
D002082:Butylamines; D014302:Trinitrobenzenesulfonic Acid; D013884:Thiosulfate Sulfurtransferase; C032462:n-butylamine