Vitamin E and glutathione are required for preservation of microsomal glutathione S-transferase from oxidative stress in microsomes.
Glutathione (GSH) inhibited lipid peroxidation induced by NADPH-BrCCl3 in vitamin E sufficient microsomes, but did not in phenobarbital (PB)-treated microsomes (containing about 60% of normal vitamin E) or in vitamin E-deficient microsomes (containing about 30% of normal vitamin E). There was a good correlation between the increased formation of CHCl3 from BrCCl3 in the presence of GSH under anaerobic conditions and the vitamin E level in the microsomes. A normal level of vitamin E in microsomes was thus very important for GSH-dependent inhibition of lipid peroxidation and for the efficient formation of CHCl3 from BrCCl3. Bromosulfophthalein (BSP) eliminated the effects of GSH on lipid peroxidation and CHCl3 formation. The apparent Km and Vmax of substrates for GSH S-transferase were changed by in vivo depletion of vitamin E in microsomes, and the Vmax/Km values were significantly reduced. The enzyme activity in microsomes was inactivated following the loss of vitamin E during in vitro lipid peroxidation, and GSH prevented the loss of vitamin E and protected the enzyme from attack by free radicals. GSH inhibited lipid peroxidation induced by NADPH-Fe2+ and the loss of GSH S-transferase activity during the peroxidation in PB-treated microsomes, but did not in the case of induction by NADPH-BrCCl3. A possible relation between the microsomal GSH S-transferase activity and defense by GSH against lipid peroxidation in microsomes is discussed.
Tampo
Y
Y
Hokkaido Institute of Pharmaceutical Sciences, Otaru, Japan.
Yonaha
M
M
D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001975:Bromotrichloromethane / Q000378:metabolism; D002725:Chloroform / Q000378:metabolism; D005978:Glutathione / Q000494:pharmacology*; D005982:Glutathione Transferase / Q000378:metabolism*; D007700:Kinetics; D015227:Lipid Peroxidation / Q000187:drug effects*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013448:Sulfobromophthalein / Q000494:pharmacology; D014810:Vitamin E / Q000494:pharmacology*
D000975:Antioxidants; D013448:Sulfobromophthalein; D014810:Vitamin E; D009249:NADP; D002725:Chloroform; D005982:Glutathione Transferase; D005978:Glutathione; D001975:Bromotrichloromethane; D010634:Phenobarbital