Inhibition of cytosolic glutathione S-transferase activity from rat liver by copper.
H(2)O(2) inactivation of particular GST isoforms has been reported, with no information regarding the overall effect of other ROS on cytosolic GST activity. The present work describes the inactivation of total cytosolic GST activity from liver rats by the oxygen radical-generating system Cu(2+)/ascorbate. We have previously shown that this system may change some enzymatic activities of thiol proteins through two mechanisms: ROS-induced oxidation and non-specific Cu(2+) binding to protein thiol groups. In the present study, we show that nanomolar Cu(2+) in the absence of ascorbate did not modify total cytosolic GST activity; the same concentrations of Cu(2+) in the presence of ascorbate, however, inhibited this activity. Micromolar Cu(2+) in either the absence or presence of ascorbate inhibited cytosolic GST activity. Kinetic studies show that GSH but no 1-chloro-2,4-dinitrobenzene prevent the inhibition on cytosolic GST induced by micromolar Cu(2+) either in the absence or presence of ascorbate. On the other hand, NEM and mersalyl acid, both thiol-alkylating agents, inhibited GST activity with differential reactivity in a dose-dependent manner. Taken together, these results suggest that an inhibitory Cu(2+)-binding effect is likely to be negligible on the overall inhibition of cytosolic GST activity observed by the Cu(2+)/ascorbate system. We discuss how modification of GST-thiol groups is related to the inhibition of cytosolic GST activity.
Letelier
M E
ME
Laboratory of Pharmacology, Department of Pharmacological and Toxicological Chemistry, Chemical and Pharmaceutical Sciences School, Universidad de Chile, Olivos 1007, Independencia, Santiago, Chile. mel@ciq.uchile.cl
Martínez
M
M
González-Lira
V
V
Faúndez
M
M
Aracena-Parks
P
P
D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology*; D001665:Binding Sites; D003300:Copper / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism*; D004137:Dinitrochlorobenzene / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors*; D007700:Kinetics; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008634:Mersalyl / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism
D000477:Alkylating Agents; D004137:Dinitrochlorobenzene; D004791:Enzyme Inhibitors; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D008634:Mersalyl; D003300:Copper; D005982:Glutathione Transferase; D001205:Ascorbic Acid