Hales B F, Neims A H
Biochem J. 1976 Nov 15;160(2):223-9. doi: 10.1042/bj1600223.
The glutathione S-transferases are a group of proteins with overlapping substrate specificities and ligand-binding capacities. This report examines certain approaches to the measurement of transferase B (ligandin) in the rat liver. The ratio of catalytic activities toward 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene gives some indication of the relative proportions of the various transferases present in 100 000 g supernatants. The fraction of catalytic activity towards 1-chloro-2,4-dinitrobenzene, due to transferase B, was best measured by immunoprecipitation with anti-(transferase B). Male rat liver exhibited three times more activity towards 1,2-dichloro-4-nitrobenzene than female tissue; however, the activities towards 1-chloro-2,4-dinitrobenzene were almost identical. By assuming a specific activity of 11 mumol/min per mg, immunoprecipitable transferase B comprised 4.5 +/- 0.2% of total protein in the 100 000 g supernatant of female rat liver, and 70% of the transferase activity towards 1-chloro-2,4-dinitrobenzene. The amount of transferase B in the 100 000 g supernatant from male rat liver is significantly lower with respect to both fraction of total protein (3.3 +/- 0.2%) and overall transferase activity towards 1-chloro-2,4-dinitrobenzene (48%). Hypophysectomy eliminated this sex difference in the hepatic concentration of glutathione S-transferase B.
谷胱甘肽S-转移酶是一组底物特异性和配体结合能力重叠的蛋白质。本报告研究了测定大鼠肝脏中转移酶B(配体结合蛋白)的某些方法。对1-氯-2,4-二硝基苯和1,2-二氯-4-硝基苯的催化活性之比可给出一些关于100000g上清液中各种转移酶相对比例的指示。由于转移酶B导致的对1-氯-2,4-二硝基苯的催化活性部分,最好通过用抗(转移酶B)进行免疫沉淀来测定。雄性大鼠肝脏对1,2-二氯-4-硝基苯的活性比雌性组织高两倍;然而,对1-氯-2,4-二硝基苯的活性几乎相同。假设每毫克的比活性为11μmol/分钟,免疫沉淀的转移酶B在雌性大鼠肝脏100000g上清液中占总蛋白的4.5±0.2%,且占对1-氯-2,4-二硝基苯转移酶活性的70%。雄性大鼠肝脏100000g上清液中转移酶B的量在总蛋白比例(3.3±0.2%)和对1-氯-2,4-二硝基苯的总体转移酶活性(48%)方面均显著较低。垂体切除术消除了肝脏中谷胱甘肽S-转移酶B浓度的这种性别差异。