Arkblad Eva L, Tuck Simon, Pestov Nikolay B, Dmitriev Ruslan I, Kostina Maria B, Stenvall Jörgen, Tranberg Mattias, Rydström Jan
Umeå Center for Molecular Pathogenesis, Umeå University, SE-901 87 Umeå, Sweden.
Free Radic Biol Med. 2005 Jun 1;38(11):1518-25. doi: 10.1016/j.freeradbiomed.2005.02.012.
Proton-translocating mitochondrial nicotinamide nucleotide transhydrogenase (NNT) was investigated regarding its physiological role in Caenorhabditis elegans. NNT catalyzes the reduction of NADP(+) by NADH driven by the electrochemical proton gradient, Deltap, and is thus a potentially important source of mitochondrial NADPH. Mitochondrial detoxification of reactive oxygen species (ROS) by glutathione-dependent peroxidases depends on NADPH for regeneration of reduced glutathione. Transhydrogenase may therefore be directly involved in the defense against oxidative stress. nnt-1 deletion mutants of C. elegans, nnt-1(sv34), were isolated and shown to grow essentially as wild type under normal laboratory conditions, but with a strongly lowered GSH/GSSG ratio. Under conditions of oxidative stress, caused by the superoxide-generating agent methyl viologen, growth of worms lacking nnt-1 activity was severely impaired. A similar result was obtained by using RNAi. Reintroducing nnt-1 in the nnt-1(sv34) knockout mutant led to a partial rescue of growth under oxidative stress conditions. These results provide evidence for the first time that nnt-1 is important in the defense against mitochondrial oxidative stress.
对质子转运线粒体烟酰胺核苷酸转氢酶(NNT)在秀丽隐杆线虫中的生理作用进行了研究。NNT催化由电化学质子梯度Δp驱动的NADH对NADP(+)的还原,因此是线粒体NADPH的一个潜在重要来源。谷胱甘肽依赖性过氧化物酶对活性氧(ROS)的线粒体解毒作用依赖于NADPH来再生还原型谷胱甘肽。因此,转氢酶可能直接参与抗氧化应激防御。分离出秀丽隐杆线虫的nnt-1缺失突变体nnt-1(sv34),并表明其在正常实验室条件下基本与野生型一样生长,但谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)比值大幅降低。在由超氧化物产生剂甲基紫精引起的氧化应激条件下,缺乏nnt-1活性的线虫生长严重受损。使用RNA干扰(RNAi)也得到了类似结果。在nnt-1(sv34)基因敲除突变体中重新引入nnt-1导致在氧化应激条件下生长得到部分挽救。这些结果首次证明nnt-1在防御线粒体氧化应激中很重要。