The role of human glutathione S-transferases hGSTA1-1 and hGSTA2-2 in protection against oxidative stress.
In order to elucidate the protective role of glutathione S-transferases (GSTs) against oxidative stress, we have investigated the kinetic properties of the human alpha-class GSTs, hGSTA1-1 and hGSTA2-2, toward physiologically relevant hydroperoxides and have studied the role of these enzymes in glutathione (GSH)-dependent reduction of these hydroperoxides in human liver. We have cloned hGSTA1-1 and hGSTA2-2 from a human lung cDNA library and expressed both in Escherichia coli. Both isozymes had remarkably high peroxidase activity toward fatty acid hydroperoxides, phospholipid hydroperoxides, and cumene hydroperoxide. In general, the activity of hGSTA2-2 was higher than that of hGSTA1-1 toward these substrates. For example, the catalytic efficiency (kcat/Km) of hGSTA1-1 for phosphatidylcholine (PC) hydroperoxide and phosphatidylethanolamine (PE) hydroperoxide was found to be 181.3 and 199.6 s-1 mM-1, respectively, while the catalytic efficiency of hGSTA2-2 for PC-hydroperoxide and PE-hydroperoxide was 317.5 and 353 s-1 mM-1, respectively. Immunotitration studies with human liver extracts showed that the antibodies against human alpha-class GSTs immunoprecipitated about 55 and 75% of glutathione peroxidase (GPx) activity of human liver toward PC-hydroperoxide and cumene hydroperoxide, respectively. GPx activity was not immunoprecipitated by the same antibodies from human erythrocyte hemolysates. These results show that the alpha-class GSTs contribute a major portion of GPx activity toward lipid hydroperoxides in human liver. Our results also suggest that GSTs may be involved in the reduction of 5-hydroperoxyeicosatetraenoic acid, an important intermediate in the 5-lipoxygenase pathway.
Zhao
T
T
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas, 77555, USA.
Singhal
S S
SS
Piper
J T
JT
Cheng
J
J
Pandya
U
U
Clark-Wronski
J
J
Awasthi
S
S
Awasthi
Y C
YC
D004306:Dose-Response Relationship, Immunologic; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D015289:Leukotrienes / Q000378:metabolism; D008099:Liver / Q000201:enzymology; D018384:Oxidative Stress / Q000187:drug effects; D011233:Precipitin Tests
D015289:Leukotrienes; C025348:arachidonic acid 5-hydroperoxide; D005979:Glutathione Peroxidase; D005982:Glutathione Transferase