Lee E, Okuno S, Kariya K
Biochem Pharmacol. 1986 Jun 1;35(11):1835-9. doi: 10.1016/0006-2952(86)90300-x.
Repeated administration of propylthiouracil (PTU) resulted in an increase in glutathione (GSH) transferases activity in rat liver cytosol toward various substrates except for epoxy(p-nitrophenoxy)propane. The enzyme from rat treated with PTU showed high activity with 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid. GSH transferases were separated into five forms by CM-Sephadex C-50 column chromatography to detect which isozymes were induced by PTU treatment. Although the activity of the unretained fraction obtained from the column was slightly increased by PTU treatment, the treatment markedly elevated the activity of GSH transferase 1-1. On the other hand, the activities of GSH transferases 1-2, 3-3 and 2-2 were little changed. In addition, an analysis of GSH transferase 1-1 from CM-Sephadex C-50 using SDS-PAGE confirmed that it comprised the 1-1 homodimer. This fraction was then further purified by passing it through a hydroxylapatite column and the partially purified GSH transferase 1-1 from rats treated with PTU was found to have the same characteristics as the control, e.g. the same Km values for GSH and CDNB and the same substrate spectrum. It was concluded that PTU specifically induced GSH transferase 1-1 among the cationic transferases.
反复给予丙硫氧嘧啶(PTU)可导致大鼠肝细胞溶质中谷胱甘肽(GSH)转移酶对除环氧(对硝基苯氧基)丙烷以外的各种底物的活性增加。用PTU处理的大鼠的该酶对1-氯-2,4-二硝基苯(CDNB)和依他尼酸表现出高活性。通过CM-葡聚糖凝胶C-50柱色谱将GSH转移酶分离为五种形式,以检测哪些同工酶是由PTU处理诱导的。尽管通过PTU处理,从柱中获得的未保留部分的活性略有增加,但该处理显著提高了GSH转移酶1-1的活性。另一方面,GSH转移酶1-2、3-3和2-2的活性几乎没有变化。此外,使用SDS-PAGE对来自CM-葡聚糖凝胶C-50的GSH转移酶1-1进行分析证实,它由1-1同二聚体组成。然后将该部分通过羟基磷灰石柱进一步纯化,发现用PTU处理的大鼠的部分纯化的GSH转移酶1-1具有与对照相同的特性,例如对GSH和CDNB具有相同的Km值以及相同的底物谱。得出的结论是,PTU在阳离子转移酶中特异性诱导GSH转移酶1-1。