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本文用S-己基谷胱甘肽-琼脂糖-6B亲和层析一步纯化法分别获得电泳纯大鼠正常肝GST_S及含增生结节大鼠肝GST_S。经DE_(52)阴离子交換柱将含增生结节大鼠肝GST_S分离为三个同功酶组份,依次命名为C_(DE)A1及A2,C_(DE)占上柱总GST_S活性84.8％。等电聚焦电泳测定等电点分别为7.8、6.7及6.3。经CM_(52)阳离子交換柱获得五个同功酶组份,依次命名为A_(CM),C1,C2,C3及C4,等电点分别为7.8,7.4,7.9,8.3及8.6。A_(CM)的活性占CM_(52)柱上柱总活性的10％。SDS-PAGE电泳结果和正常大鼠肝GST_S比较,含增生结节大鼠肝GST_S同样出现Ya,Yb及Yc三条区带,而后者的氨基酸组成也与正常大鼠肝GST_S相近,但是和大鼠正常肝组织比较后者GST_S活性明显升高,以阳离子同工酶的活性为主。
In this paper, GST_S of normal liver and GST-S of rat nodules containing hyperplastic nodules were obtained by single-step purification using S-hexyl glutathione-Sepharose-6B affinity chromatography. The hepatic GST-containing rat nodules were separated into three isoenzyme components by DE 52 anion exchange column, and named as C_ (DE) A1 and A2 in turn. C_ (DE) accounted for 84.8% . Isoelectric focusing electrophoresis determination of isoelectric points were 7.8,6.7 and 6.3. The five isoenzyme components were obtained by CM 52 cation exchange column and named as A CM, C1, C2, C3 and C4, respectively. The isoelectric points were 7.8, 7.4, 7.9, 8.3 and 8.6 respectively. The activity of A_ (CM) accounted for 10% of the total activity on the CM_ (52) column. Compared with the normal rat liver GST-S, SDS-PAGE electrophoresis showed that GST-S in the liver of rats with hyperplastic nodules also showed three bands of Ya, Yb and Yc, while the amino acid composition of the latter was also similar to that of normal rat liver GST- The activity of GST_S in the normal liver tissue was significantly higher than that in the normal liver tissue.