Butera L, Feinfeld D A, Bhargava M
Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, Bronx, N.Y.
Enzyme. 1990;43(4):175-82. doi: 10.1159/000468728.
Glutathione-S-transferase (GST) isoenzymes were purified from cytosolic preparations from kidneys of male and female rats and kidney cortical specimens from 2 male and 1 female human subjects. GST isoenzyme expression was analyzed by SDS-PAGE, measurement of catalytic activities with specific substrates and determination of their subunits by ELISA and Western blotting using specific antibodies. GST from female rat kidneys showed a preponderance of subunits 3 and 4; levels of these isoenzymes were 3-4 times greater in females than in males. Levels of subunits 1 and 2 were 1.5-2 times greater in the male rat kidneys. Additional minor bands at 24 and 22 kD were observed in GST preparations from both male and female rat kidneys while a band at 25.3 kD was observed only in the male rat kidney. These bands did not react with antibodies to GST 1-1, GST 2-2 or GST 3-4. Both male and female human kidney samples contained GST isoenzymes comparable to the near-neutral (25-5 kD) and basic forms (25 kD) of GSTs found in human liver. In addition a 28-kD band was present in GST preparations from both male and female human kidneys. Additional bands at 29 and 25.2 kD were present only in male human kidneys. Both the kidney cytosol and the total GSTs prepared from female rats shared 2- to 4-fold greater activity with 1,2-dichloro-4-nitrobenzene, ethacrynic acid and trans-4-phenyl-3-buten-2-one than those from males. The measurement of specific subunit amounts by ELISA were in agreement with these results.(ABSTRACT TRUNCATED AT 250 WORDS)
谷胱甘肽 - S - 转移酶（GST）同工酶从雄性和雌性大鼠肾脏的胞质制剂以及2名男性和1名女性人类受试者的肾皮质标本中纯化得到。通过SDS - PAGE分析GST同工酶表达，用特定底物测量催化活性，并使用特异性抗体通过ELISA和蛋白质印迹法测定其亚基。雌性大鼠肾脏中的GST显示亚基3和4占优势；这些同工酶的水平在雌性中比雄性高3至4倍。亚基1和2的水平在雄性大鼠肾脏中高1.5至2倍。在雄性和雌性大鼠肾脏的GST制剂中均观察到24 kD和22 kD处的额外小条带，而仅在雄性大鼠肾脏中观察到25.3 kD处的条带。这些条带不与针对GST 1 - 1、GST 2 - 2或GST 3 - 4的抗体发生反应。男性和女性人类肾脏样本中的GST同工酶与在人类肝脏中发现的接近中性（25 - 5 kD）和碱性形式（25 kD）的GST相当。此外，在男性和女性人类肾脏的GST制剂中均存在一条28 kD的条带。29 kD和25.2 kD处的额外条带仅存在于男性人类肾脏中。雌性大鼠制备的肾胞质溶胶和总GST与1,2 - 二氯 - 4 - 硝基苯、依他尼酸和反式 - 4 - 苯基 - 3 - 丁烯 - 2 - 酮的活性比雄性大鼠的高2至4倍。通过ELISA测量特定亚基的量与这些结果一致。（摘要截断于250字）
