[Studies on preferential binding to glucocorticoid of rat liver anionic glutathione S-transferase].
A new isozyme of Glutathione-S-transferase (GST) with more acidic pI (6.7) than other forms of GST hitherto reported was isolated from rat liver cytosol by consecutive chromatographies on a DEAE cellulose column, lysyl-GSH affinity column and Sephadex G-100 column. This anionic form of GST represented approximately one third of total GST activity in rat liver cytosol. Amino acid composition, immunological reactivity, enzymatic properties, and secondary structure as measured by circular dichroism of this form were distinct from those of cationic isozymes (GST-AA, GST-B, GST-X), presently investigated. The stoichiometric ratio of high affinity site for bilirubin to GST molecule differs amongst isozymes. The anionic form of GST bound two bilirubin per molecule whereas cationic GSTs bound only one bilirubin per two subunits. The most distinguished property of anionic GST was its strong affinity for glucocorticoid. The dissociation constant of anionic GST-corticosterone complex was as low as 2.0 X 10(-8) M. Corticosterone inhibited the enzyme activity of anionic GST in a noncompetitive fashion with an apparent Ki value of 8.6 X 10(-5) M and 1.1 X 10(-6) M for 1-chloro-2.4-dinitrobenzene and GST respectively. The anionic GST-corticosterone complex bound to DNA coupled Sepharose at 25 degrees C and passed through the column at 4 degrees C. Conversely, the complex bound to a DEAE cellulose column at 4 degrees C but passed through at 25 degrees C. These properties of anionic GST are quite similar to those of glucocorticoid receptor of rat liver cytosol reported previously.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000838:Anions; D001663:Bilirubin / Q000378:metabolism; D003345:Corticosterone / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D004586:Electrophoresis; D005982:Glutathione Transferase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D000596:Amino Acids; D000838:Anions; D005982:Glutathione Transferase; D001663:Bilirubin; D003345:Corticosterone