Quantifying Reductive Carboxylation Flux of Glutamine to Lipid in a Brown Adipocyte Cell Line
Open accessAbstract
We previously reported that glutamine was a major source of carbon for de novo fatty acid synthesis in a brown adipocyte cell line. The pathway for fatty acid synthesis from glutamine may follow either of two distinct pathways after it enters the citric acid cycle. The glutaminolysis pathway follows the citric acid cycle, whereas the reductive carboxylation pathway travels in reverse of the citric acid cycle from alpha-ketoglutarate to citrate. To quantify fluxes in these pathways we incubated brown adipocyte cells in [U-(13)C]glutamine or [5-(13)C]glutamine and analyzed the mass isotopomer distribution of key metabolites using models that fit the isotopomer distribution to fluxes. We also investigated inhibitors of NADP-dependent isocitrate dehydrogenase and mitochondrial citrate export. The results indicated that one third of glutamine entering the citric acid cycle travels to citrate via reductive carboxylation while the remainder is oxidized through succinate. The reductive carboxylation flux accounted for 90% of all flux of glutamine to lipid. The inhibitor studies were compatible with reductive carboxylation flux through mitochondrial isocitrate dehydrogenase. Total cell citrate and alpha-ketoglutarate were near isotopic equilibrium as expected if rapid cycling exists between these compounds involving the mitochondrial membrane NAD/NADP transhydrogenase. Taken together, these studies demonstrate a new role for glutamine as a lipogenic precursor and propose an alternative to the glutaminolysis pathway where flux of glutamine to lipogenic acetyl-CoA occurs via reductive carboxylation. These findings were enabled by a new modeling tool and software implementation (Metran) for global flux estimation.
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IDH-dependent reductive carboxylation has been investigated as a way to produce citrate and fatty acids from α-ketoglutarate derived from glutamine.
“has been investigated since then as a way to produce citrate and fatty acids from α-ketoglutarate derived from glutamine (Holleran et al., 1995; Ward et al., 2010; Yoo et al., 2008)”
From Metabolic Reprogramming: A Cancer Hallmark Even Warburg Did Not Anticipate (2012) · cites this paper as Yoo et al., 2008
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- Authors
- Yoo, H., Antoniewicz, M. R., Stephanopoulos, G., & Kelleher, J. K.
- Affiliations
- Massachusetts Institute of Technology (US)
- Corresponding author
- Yoo, Hyuntae
- Antoniewicz, Maciek R.
- Kelleher, Joanne K.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2008-07
- Volume
- 283
- Issue
- 30
- Pages
- 20621-20627
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC2475696
- OpenAlex
- W2033471108
- Cited by
- 301
- References
- 26
- Citations by year
- 2026: 5 · 2025: 7 · 2024: 14 · 2023: 19 · 2022: 19 · 2021: 12 · 2020: 21 · 2019: 21 · 2018: 14 · 2017: 16 · 2016: 23 · 2015: 20 · 2014: 27 · 2013: 23 · 2012: 23
- FWCI
- 4.10
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Joslin Diabetes Center
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases (DK 075850, R01DK075850)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- reductive carboxylation, glutamine, brown adipocytes, de novo lipogenesis, TCA cycle, isocitrate dehydrogenase, alpha-ketoglutarate, mass isotopomer distributions, flux analysis, lipogenesis, acetyl-CoA
- MeSH terms
- Acetyl Coenzyme A, Animals, Carboxylic Acids, Cell Differentiation, Cell Line, Citric Acid Cycle, Fatty Acids, Glutamine, Isocitrate Dehydrogenase, Ketoglutaric Acids, Lipids, Mitochondria, Oxalates, Mice, Adipocytes, Brown
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