Glutamine metabolism in AS-30D hepatoma cells. Evidence for its conversion into lipids via reductive carboxylation
Abstract
A study was undertaken to assess the role of a physiological concentration of glutamine in AS-30D cell metabolism. Flux of 14C-glutamine to 14CO2 and of 14C-acetate to glutamate was detected indicating reversible flux between glutamate and TCA cycle alpha-ketoglutarate. These fluxes were transaminase dependent. A flux analysis was compared using data from three tracers that label alpha-ketoglutarate carbon 5, [2-14C]glucose, [1-14C]acetate and [5-14C]glutamine. The analysis indicated that the probability of flux of TCA cycle alpha-ketoglutarate to glutamate was, at minimum, only slightly less than the probability of flux of alpha-ketoglutarate through alpha-ketoglutarate dehydrogenase. The apparent Km for oxidative flux of [14C]glutamine to 14CO2, 0.07 mM, indicated that this flux was at a maximal rate at physiological, 0.75 mM, glutamine. Although oxidative flux through alpha-ketoglutrate dehydrogenase was the major fate of glutamine, flux of glutamine to lipid via reductive carboxylation of alpha-ketoglutarate was demonstrated by measuring incorporation of [5-14C]glutamine into 14C-lipid. In media containing glucose (6 mM), and glutamine (0.75 mM) 47 per cent of the lipid synthesized from substrates in the media was derived from glutamine via reductive carboxylation and 49 per cent from glucose. These findings of nearly equal fluxes suggest that lipogenesis via reductive carboxylation may be an important role of glutamine in hepatoma cells.
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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 Holleran et al., 1995
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- Authors
- Holleran, A. L., Briscoe, D. A., Fiskum, G., & Kelleher, J. K.
- Affiliations
- George Washington University (US)
- ORCID
- Fiskum, Gary
- Venue
- Molecular and Cellular Biochemistry
- Published
- 1995-11
- Volume
- 152
- Issue
- 2
- Pages
- 95-101
- ISSN
- 0300-8177 (ISSN-L)
- 1573-4919
- OpenAlex
- W2005093036
- Cited by
- 53
- References
- 23
- Citations by year
- 2026: 2 · 2024: 2 · 2023: 1 · 2022: 2 · 2021: 5 · 2020: 1 · 2019: 5 · 2017: 2 · 2016: 1 · 2015: 8 · 2014: 2 · 2013: 5 · 2012: 4
- FWCI
- 0.16
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Molecular and Cellular Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01gm033536-08, 5r01ca032946-08)
- National Cancer Institute (CA-32946)
- National Institute of General Medical Sciences (GM33536)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
- Keywords
- glutamine metabolism, reductive carboxylation, lipogenesis, alpha-ketoglutarate, glutamate, TCA cycle, alpha-ketoglutarate dehydrogenase, transamination, glucose metabolism, flux analysis
- MeSH terms
- Animals, Carboxylic Acids, Citric Acid Cycle, Glucose, Glutamine, Ketoglutaric Acids, Liver Neoplasms, Experimental, Oxidation-Reduction, Tumor Cells, Cultured, Glutamic Acid, Lipid Metabolism, Rats
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