Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio.
Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio.
复制标题
DOI：
10.1126/science.aad4017
复制
发表时间：
2016-04-08
期刊：
影响因子：
--
通讯作者：
Mootha VK
中科院分区：
文献类型：
--
作者：
Titov DV;Cracan V;Goodman RP;Peng J;Grabarek Z;Mootha VK
电子传递链（ETC）活性的下降与许多人类疾病有关。虽然线粒体ATP产生减少被认为是病理学的一个来源，但氧化型烟酰胺腺嘌呤二核苷酸（NAD+）与其还原型（NADH）的量之比的相关减少的贡献尚不清楚。我们使用了一种产水的L.因此，本发明提供了一种用于在人细胞中诱导NAD+/NADH比率的隔室特异性增加的基因工具，即LbNOX。我们使用LbNOX来证明关键代谢通量、代谢产物生成和信号传导对细胞溶质或线粒体NAD+/NADH比率的依赖性。LbNOX在细胞质或线粒体中的表达改善了由受损的ETC引起的增殖和代谢缺陷。结果强调了还原应激在线粒体发病机制中的作用，并证明了靶向LbNOX用于直接的、隔室特异性的氧化还原状态操纵的实用性。我们开发了一种用于提高NAD+/NADH比率的遗传编码工具，并表明它可以补充人类细胞中受损的电子传递链。
A decline in electron transport chain (ETC) activity is associated with many human diseases. Although diminished mitochondrial ATP production is recognized as a source of pathology, the contribution of the associated reduction in the ratio of the amount of oxidized nicotinamide adenine dinucleotide (NAD+) to that of its reduced form (NADH) is less clear. We used a water-forming NADH oxidase from L. brevis (LbNOX) as a genetic tool for inducing a compartment-specific increase of the NAD+/NADH ratio in human cells. We used LbNOX to demonstrate the dependence of key metabolic fluxes, gluconeogenesis, and signaling on the cytosolic or mitochondrial NAD+/NADH ratios. Expression of LbNOX in the cytosol or mitochondria ameliorated proliferative and metabolic defects caused by an impaired ETC. The results underscore the role of reductive stress in mitochondrial pathogenesis and demonstrate the utility of targeted LbNOX for direct, compartment-specific manipulation of redox state. We developed a genetically encoded tool for raising NAD+/NADH ratios and showed it can complement an impaired electron transport chain in human cells.