Steisslinger H W, Pfleiderer G
Institut für Organische Chemie, Biochemie und Isotopenforschung, Universität, Stuttgart, BRD.
Enzyme. 1988;40(1):1-6. doi: 10.1159/000469133.
Glutathione S-transferase (GSH-transferase) was purified from human placenta and kidney by affinity chromatography on S-glutathione-carbamidomethyl-epsilon-aminolysyl-Sepharose CL 4B and gel filtration chromatography on Sephades G-75. Electrophoretically pure enzyme with the specific activities of 50.7 and 55.9 U/mg, respectively, were obtained. In addition to the known acidic isoenzyme from human placenta (isoelectric point, pI, 4.5), we describe here for the first time the presence of 6 basic forms with pI values between 8.0 and 9.0. The kidney GSH-transferase contained 2 acidic forms with isoelectric points at 4.6 and 4.65, and 6 basic forms with pI values between 8.7 and 9.4. The basic and acidic isoenzymes from placenta were separated by ion exchange chromatography on Sephadex DEAE A-25. The acidic form accounted for 36% of the total GSH-transferase activity from placenta. Antibodies against the kidney enzyme were raised in rabbit. Total cross-reactivity of placental GSH-transferase with antikidney-GSH-transferase antibodies was obtained, suggesting that the kidney and placental enzymes are immunologically closely related.
通过在S-谷胱甘肽-氨甲酰甲基-ε-氨基赖氨酸-琼脂糖凝胶CL 4B上进行亲和层析以及在Sephades G-75上进行凝胶过滤层析，从人胎盘和肾脏中纯化出谷胱甘肽S-转移酶（GSH-转移酶）。分别获得了比活性为50.7和55.9 U/mg的电泳纯酶。除了已知的人胎盘酸性同工酶（等电点，pI，4.5）外，我们首次在此描述了存在6种pI值在8.0至9.0之间的碱性形式。肾脏GSH-转移酶含有2种等电点分别为4.6和4.65的酸性形式，以及6种pI值在8.7至9.4之间的碱性形式。胎盘的碱性和酸性同工酶通过在Sephadex DEAE A-25上进行离子交换层析分离。酸性形式占胎盘总GSH-转移酶活性的36%。用兔制备了针对肾脏酶的抗体。获得了胎盘GSH-转移酶与抗肾脏GSH-转移酶抗体的完全交叉反应性，表明肾脏和胎盘的酶在免疫学上密切相关。
