In fish, glutathione S-transferases are very important in the detoxification of water-borne pollutants. Carp, which often live and thrive under extreme conditions of water pollution, have been recommended by the European Community as an ideal species to test for the effect of various chemical substances often found in the aqueous environment. In this paper we describe the purification and kinetic characterization of the three major forms of glutathione S-transferase in carp liver cytosol. The main step for purification was affinity chromatography on S-hexylglutathione-Sepharose 6B and the isoenzymes have been separated by isoelectric focusing at pH 7.7, 7.3, 7.1. Values of Km and kcat/Km for both reduced glutathione and 1-chloro-2,4-dinitrobenzene as substrates have been determined, and the isoenzymes characterized with several substrates and inhibitors.
Characterizazion of the major glutathione transferases in carp liver
ELIA, Antonia Concetta;MANTILACCI, Luciana;TATICCHI, Maria Illuminata;
1999
Abstract
In fish, glutathione S-transferases are very important in the detoxification of water-borne pollutants. Carp, which often live and thrive under extreme conditions of water pollution, have been recommended by the European Community as an ideal species to test for the effect of various chemical substances often found in the aqueous environment. In this paper we describe the purification and kinetic characterization of the three major forms of glutathione S-transferase in carp liver cytosol. The main step for purification was affinity chromatography on S-hexylglutathione-Sepharose 6B and the isoenzymes have been separated by isoelectric focusing at pH 7.7, 7.3, 7.1. Values of Km and kcat/Km for both reduced glutathione and 1-chloro-2,4-dinitrobenzene as substrates have been determined, and the isoenzymes characterized with several substrates and inhibitors.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
