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Differential effects of insulin signaling on individual carbon fluxes for fatty acid synthesis in brown adipocytes

Author(s)
Yoo, Hyuntae; Antoniewicz, Maciek; Kelleher, Joanne K.; Stephanopoulos, Gregory
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Abstract
Considering the major role of insulin signaling on fatty acid synthesis via stimulation of lipogenic enzymes, differential effects of insulin signaling on individual carbon fluxes for fatty acid synthesis have been investigated by comparing the individual lipogenic fluxes in WT and IRS-1 knockout (IRS-1 KO) brown adipocytes. Results from experiments on WT and IRS-1 KO cells incubated with [5-¹³C] glutamine were consistent with the existence of reductive carboxylation pathway. Analysis of isotopomer distribution of nine metabolites related to the lipogenic routes from glucose and glutamine in IRS-1 KO cells using [U-¹³C] glutamine as compared to that in WT cells indicated that flux through reductive carboxylation pathway was diminished while flux through conventional TCA cycle was stimulated due to absence of insulin signaling in IRS-1 KO cells. This observation was confirmed by quantitative estimation of individual lipogenic fluxes in IRS-1 KO cells and their comparison with fluxes in WT cells. Thus, these results suggest that glutamine’s substantial contribution to fatty acid synthesis can be directly manipulated by controlling the flux through reductive carboxylation of alpha-ketoglutarate to citrate using hormone (insulin).
Date issued
2005-01
URI
http://hdl.handle.net/1721.1/7478
Series/Report no.
Molecular Engineering of Biological and Chemical Systems (MEBCS);
Keywords
brown adipocyte, fatty acid synthesis, insulin signaling, reductive carboxylation pathway, 5-13C, U-13C

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