Mishra Rajnikant, Shukla S P
Department of Zoology, Banaras Hindu University, 221005, Varanasi, India.
Ecotoxicol Environ Saf. 2003 Nov;56(3):425-33. doi: 10.1016/s0147-6513(03)00006-x.
The effects of a sublethal concentration of an organochlorine pesticide endosulfan were investigated on crude and purified forms of cytoplasmic malate dehydrogenase (cMDH) and mitochondrial malate dehydrogenase (mMDH) in the muscle of the freshwater catfish Clarias batrachus. Endosulfan treatment reduced significantly the activity and the specific activity of cMDH and mMDH but had no effect on total cytoplasmic and mitochondrial protein contents. This indicates a decline in the efficiency of aerobic energy metabolism in endosulfan-treated fish. The polyacrylamide gel electrophoresis showed two distinct isoforms (C1 and C2) of cMDH and only one form of mMDH. The inhibition produced by endosulfan in vivo was of mixed noncompetitive/uncompetitive type for crude as well as purified cMDH and mMDH. However, in vitro inhibition was of simple noncompetitive and mixed competitive/noncompetitive type for purified cMDH and mMDH, respectively. Citrate was found to be an uncompetitive inhibitor of cMDH and mixed noncompetitive/uncompetitve inhibitor of mMDH. The inhibitory patterns were modulated by endosulfan. These results demonstrate inhibitory effects of endosulfan on skeletal muscle MDH of the freshwater catfish Clarias batrachus and inhibition effects are mediated through enzyme/substrate/endosulfan complexing.
研究了亚致死浓度的有机氯农药硫丹对淡水鲶鱼(胡子鲶)肌肉中粗提和纯化形式的细胞质苹果酸脱氢酶(cMDH)和线粒体苹果酸脱氢酶(mMDH)的影响。硫丹处理显著降低了cMDH和mMDH的活性及比活性,但对细胞质和线粒体总蛋白含量没有影响。这表明硫丹处理的鱼有氧能量代谢效率下降。聚丙烯酰胺凝胶电泳显示cMDH有两种不同的同工型(C1和C2),而mMDH只有一种形式。硫丹在体内对粗提和纯化的cMDH及mMDH产生的抑制作用为混合型非竞争性/非竞争性抑制。然而,在体外,对纯化的cMDH和mMDH的抑制作用分别为单纯非竞争性抑制和混合型竞争性/非竞争性抑制。发现柠檬酸是cMDH的非竞争性抑制剂和mMDH的混合型非竞争性/非竞争性抑制剂。硫丹调节了抑制模式。这些结果证明了硫丹对淡水鲶鱼骨骼肌MDH的抑制作用,且抑制作用是通过酶/底物/硫丹络合介导的。