Dog liver glutathione S-transferase and its strong immunoreactivity with rat transferase-P(7-7).
Dog liver cytosolic glutathione S-transferases (GSTs) were investigated to characterize their properties in comparison with rat liver transferases. Dog liver GSTs after the glutathione affinity column chromatography showed three subunit bands on SDS-polyacrylamide gel electrophoresis. These three subunits, designated as Yd1 (mol.wt 26,000), Yd2 (mol.wt 27,000) and Yd3 (mol.wt 28,000), were distinctly different from rat liver GST subunits, i.e. Ya(1) (mol.wt 26,500), Yb1(3)/Yb2(4) (mol.wt 27,500) and Yc(2) (mol.wt 28,500). Western blot analysis revealed that Yd1, Yd2 and Yd3 were immunoreacted with anti-rat GST 7-7, 1-1 and 3-3 antibodies, respectively. Four transferase activity fractions, I (pH greater than 7.63), II (pH 6.92), III (pH 5.80) and IV (pH 5.65), were obtained from affinity purified GSTs by chromatofocusing. Each fraction exhibited a characteristic substrate specificity. GST-II, III and IV were all strongly immunoreacted with anti-rat GST 7-7 antibody by immunoblotting, thus suggesting the occurrence of the heterogeneity of transferases immunologically related to rat GST subunit 7 in dog liver. Immunohistochemical examination showed that transferases immunoreacted with anti-GST 7-7 antibody have diffusely distributed throughout the lobule, while enzymes related to subunit 3 have been localized in a narrow range of cells around the central vein. These data suggest that GSTs immunologically associated with rat transferase subunit 7 may be major forms in dog liver.
Igarashi
T
T
Department of Biochemical Pharmacology and Biotoxicology, Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Nanba
E
E
Sagami
F
F
Tsukidate
K
K
Fukuda
T
T
Horie
T
T
Satoh
T
T
Kitagawa
H
H
D000818:Animals; D015153:Blotting, Western; D003600:Cytosol / Q000201:enzymology; D004285:Dogs / Q000378:metabolism*; D005982:Glutathione Transferase / Q000276:immunology* / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007526:Isoelectric Point; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D051381:Rats; D013379:Substrate Specificity
D046911:Macromolecular Substances; D005982:Glutathione Transferase