TY - JOUR
T1 - Cytosolic and mitochondrial NADPH fluxes are independently regulated
AU - Niu, Xiangfeng
AU - Stancliffe, Ethan
AU - Gelman, Susan J.
AU - Wang, Lingjue
AU - Schwaiger-Haber, Michaela
AU - Rowles, Joe L.
AU - Shriver, Leah P.
AU - Patti, Gary J.
N1 - Funding Information:
This work was supported by funding from National Institutes of Health grant nos. R35ES028365 (G.J.P.) and R24OD024624 (G.J.P.).
Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2023/7
Y1 - 2023/7
N2 - 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. [Figure not available: see fulltext.].
AB - 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. [Figure not available: see fulltext.].
UR - http://www.scopus.com/inward/record.url?scp=85150937808&partnerID=8YFLogxK
U2 - 10.1038/s41589-023-01283-9
DO - 10.1038/s41589-023-01283-9
M3 - Article
C2 - 36973440
AN - SCOPUS:85150937808
SN - 1552-4450
VL - 19
SP - 837
EP - 845
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 7
ER -