Cited by 563Redox biology and oxidative stressCancer, Hypoxia, and MetabolismCancer and biochemical research

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Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells

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Lewis, Caroline A., Parker, Seth J., Fiske, Brian P., McCloskey, Douglas, Gui, Dan Y., Green, Courtney R., Vokes, Natalie I., Feist, Adam M., Vander Heiden, Matthew G., Metallo, Christian M.·2014·Molecular Cell

DOI 10.1016/j.molcel.2014.05.008PMC PMC4106038

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Abstract

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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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: Lewis, Caroline A.
Parker, Seth J.
Fiske, Brian P.
McCloskey, Douglas
Gui, Dan Y.
Green, Courtney R.
Vokes, Natalie I.
Feist, Adam M.
Metallo, Christian M.
Venue: Molecular Cell
Publisher: Elsevier BV
Published: 2014-07
Volume: 55
Issue: 2
Pages: 253-263
ISSN: 1097-2765 (ISSN-L)
DOI: 10.1016/j.molcel.2014.05.008
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: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/open-access/userlicense/1.0/ (vor, 381 d)
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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Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells

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