Cytosolic and mitochondrial NADPH fluxes are independently regulated.
Cytosolic and mitochondrial NADPH fluxes are independently regulated.
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细胞质和线粒体 NADPH 通量是独立调节的。
DOI：
10.1038/s41589-023-01283-9
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发表时间：
2023
影响因子：
14.8
通讯作者：
Patti,GaryJ
中科院分区：
文献类型：
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作者：
Niu,Xiangfeng;Stancliffe,Ethan;Gelman,SusanJ;Wang,Lingjue;Schwaiger-Haber,Michaela;Rowles3rd,JoeL;Shriver,LeahP;Patti,GaryJ
尽管烟酰胺腺嘌呤二核苷酸磷酸(NADPH)在细胞质和线粒体中产生和消费，但由于技术限制，各隔室NADPH通量之间的关系很难评估。在这里，我们介绍了一种解决胞质和线粒体NADPH通量的方法，该方法依赖于从葡萄糖到胞浆或线粒体中的脯氨酸生物合成代谢产物的示踪。我们通过使用异柠檬酸脱氢酶突变、化疗药物或遗传编码的NADPH氧化酶，在细胞的胞浆或线粒体中引入了NADPH挑战。我们发现，胞质挑战影响了胞质中的NADPH通量，但不影响线粒体中的NADPH通量，反之亦然。这项工作强调了使用Pro标记作为报告系统来研究分区代谢的价值，并揭示了细胞胞浆和线粒体位置的NADPH动态平衡是独立调节的，没有证据表明NADPH穿梭活动。
Although nicotinamide adenine dinucleotide phosphate (NADPH) is produced and consumed in both the cytosol and mitochondria, the relationship between NADPH fluxes in each compartment has been difficult to assess due to technological limitations. Here we introduce an approach to resolve cytosolic and mitochondrial NADPH fluxes that relies on tracing deuterium from glucose to metabolites of proline biosynthesis localized to either the cytosol or mitochondria. We introduced NADPH challenges in either the cytosol or mitochondria of cells by using isocitrate dehydrogenase mutations, administering chemotherapeutics or with genetically encoded NADPH oxidase. We found that cytosolic challenges influenced NADPH fluxes in the cytosol but not NADPH fluxes in mitochondria, and vice versa. This work highlights the value of using proline labeling as a reporter system to study compartmentalized metabolism and reveals that NADPH homeostasis in the cytosolic and mitochondrial locations of a cell are independently regulated, with no evidence for NADPH shuttle activity.