Molecular characterisation of a NADH ubiquinone oxidoreductase subunit 5 from Schistosoma mansoni and inhibition of mitochondrial respiratory chain function by testosterone
Molecular characterisation of a NADH ubiquinone oxidoreductase subunit 5 from Schistosoma mansoni and inhibition of mitochondrial respiratory chain function by testosterone
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DOI：
10.1023/a:1007057903390
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发表时间：
1999-12-01
影响因子：
4.3
通讯作者：
Rumjanek, FD
中科院分区：
文献类型：
--
作者：
Fantappié, MR;Galina, A;Rumjanek, FD
通过使用人类雄激素受体 (hAR) cDNA 探针筛选曼氏血吸虫 cDNA 文库，分离出编码人类寄生虫曼氏血吸虫线粒体酶 NADH-泛醌氧化还原酶亚基 5 (SmND5) 的互补 DNA。确定了SmND5的完整核苷酸序列和推导的氨基酸序列。 Southern印迹分析揭示了SmND5的单拷贝基因的存在，并且通过RT-PCR，表明SmND5发生了性别和阶段特异性表达。为了建立线粒体酶和雄激素受体之间的功能关系，使用成人血吸虫和牛心亚软骨颗粒将睾酮的作用与经典呼吸链抑制剂的作用进行比较。睾酮的生理浓度能够抑制线粒体膜上质子梯度的维持以及 ATP 合成。该类固醇被发现对幼虫有细胞毒性，但对成虫没有细胞毒性。提出了一个模型来解释在实验性慢性感染中观察到的体内睾酮相关的蠕虫负荷差异。
Complementary DNA, encoding the mitochondrial enzyme NADH-ubiquinone oxidoreductase subunit 5 (SmND5) of the human parasite Schistosoma mansoni was isolated by screening a S. mansoni cDNA library with a human androgen receptor (hAR) cDNA probe. The complete nucleotide and deduced aminoacid sequences of SmND5 were determined. Southern blot analysis revealed the occurrence of a single copy gene for SmND5 and by means of RT-PCR, it was shown that sex- and stage-specific expression of SmND5 occurred. In order to establish a functional relationship between the mitochondrial enzyme and the androgen receptor, the effects of testosterone were compared to those of classical respiratory chain inhibitors, using adult schistosome and beef heart submitochondrial particles. Physiological concentrations of testosterone were able to inhibit the maintenance of proton gradient across the mitochondrial membranes, as well as ATP synthesis. The steroid was found to be cytotoxic to the larvae, but not to adult schistosomes. A model is proposed to explain the observed in vivo testosterone-related differences in worm burdens, in experimental chronic infections.