J. Lipid Res.
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A more recent version of this article appeared on July 1, 2004

Papers In Press, published online ahead of print April 21, 2004
J. Lipid Res., doi:10.1194/jlr.M400031-JLR200
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Submitted on January 27, 2004
Revised on April 8, 2004
Accepted on April 12, 2004

Quantifying carbon sources for de novo lipogenesis in wild-type and IRS-1 knockout brown adipocytes

Hyuntae Yoo, Gregory Stephanopoulos, and Joanne K. Kelleher

Chemical Engineering, MIT, Cambridge, MA 02139

Corresponding Author: jkk{at}mit.edu

Studies were conducted to evaluate the flux of various carbon sources to lipogenesis during brown adipocyte differentiation. 13C-labeling and isotopomer spectral analysis quantified the contribution of metabolites to de novo lipogenesis in wild-type (WT) and IRS-1 knockout (KO) brown adipocytes. Both glucose and glutamine provided substantial fractions of the lipogenic acetyl CoA for both WT and KO cells in standard media, together contributing 60%. Adding acetoacetate (10 mM) to the medium resulted in a large flux of acetoacetate to lipid representing 70% of the lipogenic acetyl CoA and decreasing the contribution of glucose plus glutamine to 30%. For WT cells, the fractional synthesis of new fatty acids during 4-day differentiation was 80% of the total. Similarly, 80% of the lipidic glycerol was derived from glucose in the medium; glutamine was not a precursor for glycerol. When glutamine was removed from the medium, the contribution of glucose to fatty acid synthesis doubled, replacing most of the contribution of glutamine and maintaining total lipogenesis. Conversely, removal of glucose dramatically decreased lipogenesis. These results indicate that glucose’s distinct role in lipid synthesis during differentiation cannot be replaced by other carbon sources, consistent with the role for glucose supplying NADPH and/or glycerol for triglyceride synthesis.


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H. Yoo, M. R. Antoniewicz, G. Stephanopoulos, and J. K. Kelleher
Quantifying Reductive Carboxylation Flux of Glutamine to Lipid in a Brown Adipocyte Cell Line
J. Biol. Chem., July 25, 2008; 283(30): 20621 - 20627.
[Abstract] [Full Text] [PDF]


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