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Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells
Open accessAbstract
Eukaryotic cells compartmentalize biochemical processes in different organelles, often relying on metabolic cycles to shuttle reducing equivalents across intracellular membranes. NADPH serves as the electron carrier for the maintenance of redox homeostasis and reductive biosynthesis, with separate cytosolic and mitochondrial pools providing reducing power in each respective location. This cellular organization is critical for numerous functions but complicates analysis of metabolic pathways using available methods. Here we develop an approach to resolve NADP(H)-dependent pathways present within both the cytosol and the mitochondria. By tracing hydrogen in compartmentalized reactions that use NADPH as a cofactor, including the production of 2-hydroxyglutarate by mutant isocitrate dehydrogenase enzymes, we can observe metabolic pathway activity in these distinct cellular compartments. Using this system we determine the direction of serine/glycine interconversion within the mitochondria and cytosol, highlighting the ability of this approach to resolve compartmentalized reactions in intact cells.
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“Serine can also contribute to NADPH production via the folate cycle, which serves to maintain redox homeostasis and support anabolic reactions (Fan et al., 2014; Lewis et al., 2014; Ye et al., 2014).”
From Targeting Metabolism for Cancer Therapy (2017) · cites this paper as Lewis et al., 2014
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Metadata
- Authors
- Lewis, C. A., Parker, S. J., Fiske, B. P., McCloskey, D., Gui, D. Y., Green, C. R., Vokes, N. I., Feist, A. M., Vander Heiden, M. G., & Metallo, C. M.
- Affiliations
- Koch Institute for Integrative Cancer Research At MIT (US)
- University of California San Diego (US)
- Corresponding author
- Metallo, Christian M.
- ORCID
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2014-07
- Volume
- 55
- Issue
- 2
- Pages
- 253-263
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC4106038
- OpenAlex
- W2031022790
- Cited by
- 562
- References
- 54
- Citations by year
- 2026: 22 · 2025: 26 · 2024: 47 · 2023: 48 · 2022: 56 · 2021: 58 · 2020: 66 · 2019: 54 · 2018: 45 · 2017: 50 · 2016: 47 · 2015: 28 · 2014: 15
- FWCI
- 19.12
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-nd)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- U.S. Department of Defense (W81XWH, W81XWH-13)
- Damon Runyon Cancer Research Foundation
- Cancer Research Coordinating Committee
- Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
- National Institutes of Health (W81XWH, CA121852)
- National Cancer Institute (U54-CA121852-09, U54CA121852, R01CA168653, R01CA188652, P30CA147882)
- National Institute of General Medical Sciences (T32GM007753, R01GM132106, T32GM007287)
- National Institute of Biomedical Imaging and Bioengineering (T32EB009380)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer and biochemical research
- Keywords
- NADPH metabolism, compartmentalized metabolism, mitochondrial metabolism, cytosolic metabolism, 2-hydroxyglutarate, isocitrate dehydrogenase, redox homeostasis, reducing power, metabolic pathway activity, mammalian cells
- MeSH terms
- Cytosol, Glucose, Glycine, Humans, Isocitrate Dehydrogenase, Mitochondria, NADP, Serine, Cell Line, Tumor, Metabolic Flux Analysis
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