Purification and characterization of glutathione S-transferases from guinea pig liver.
Four types of glutathione S-transferase were purified to homogeneity from guinea pig liver by DEAE-cellulose, Sephadex G-75, CM-cellulose, and affinity chromatography. These isozymes were named a, b, c, and d based on the reverse order of elution from a CM-cellulose column, and had specific activities of 89.6, 92.2, 99.0, and 44.0 units/mg, respectively, when assayed with 1 mM each of 1-chloro-2,4-dinitrobenzene and reduced glutathione. All four transferases of guinea pig liver were homodimers. The transferases b, c, and d had a similar molecular weight of 50,000 and their subunit sizes were 25,000, but the corresponding values for transferase a were 45,000 and 23,500, respectively. Transferase a was notably different in the activities towards organic hydroperoxides and 1,2-dichloro-4-nitrobenzene from the other isozymes. Transferases a and b, the major forms in guinea pig liver, were studied with respect to their biochemical properties, including kinetic parameters, absorption and fluorescence spectra, and bilirubin binding. Glutathione peroxidase activity of the transferase a was about 100 times higher than that of other isozymes. In guinea pig liver, it is estimated that transferase a is the major glutathione peroxidase, accounting for about 75% of the total organic hydroperoxide reduction.
Oshino
R
R
Laboratory of Biochemistry, Kobe Yamate Women's Junior College, Hyogo.
Kamei
K
K
Nishioka
M
M
Shin
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001663:Bilirubin / Q000378:metabolism; D002645:Chickens; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000302:isolation & purification*; D006168:Guinea Pigs; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D000596:Amino Acids; D005979:Glutathione Peroxidase; D005982:Glutathione Transferase; D001663:Bilirubin