NDLI: Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line.

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Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line.

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Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Antoniewicz, Maciek R. Stephanopoulos, Gregory Yoo, Hyuntae Kelleher, Joanne K.
Description	Author Affiliation: Yoo H ( Department of Chemistry and Department of Chemical Engineering, Bioinformatics and Metabolic Engineering Laboratory, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.)
Abstract	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.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	30
Volume Number	283
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2008-07-25
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Adipocytes, Brown Cytology Carboxylic Acids Chemistry Glutamine Lipids Acetyl Coenzyme A Animals Cell Differentiation Cell Line Citric Acid Cycle Fatty Acids Isocitrate Dehydrogenase Ketoglutaric Acids Mice Mitochondria Enzymology Oxalates Research Support, N.I.H., Extramural Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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