| Reference |
Fan J, Kamphorst JJ, Mathew R, Chung MK, White E, Shlomi T, Rabinowitz JD. Glutamine-driven oxidative phosphorylation is a major ATP source in transformed mammalian cells in both normoxia and hypoxia. Mol Syst Biol. 2013 Dec 3 9: 712. doi: 10.1038/msb.2013.65. Supplementary information p.11 table 5PubMed ID24301801
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| Method |
"To infer intracellular metabolic fluxes, [researchers] constructed a metabolic
network model of glycolysis and TCA cycle (Supplementary Table 3 BNID 110689)
and applied metabolic flux analysis (MFA) to identify a flux
distribution that optimally fits the following experimental data sets
(Supplementary Figures 1–4 and Supplementary Table 5): (i)
measured uptake and secretion rates of glucose, glutamine, lactate,
pyruvate, and alanine (ii) measured oxygen consumption rate used by
oxidative phosphorylation (iii) steady-state isotopic labeling pattern
of pyruvate, a-ketoglutarate, citrate, malate, and cytosolic acetyl-CoA,
where the labeling pattern of cytosolic acetyl-CoA was inferred based
on the steady-state labeling pattern of fatty acids via isotopomer
spectral analysis (ISA)(Kharroubi et al, 1992) (iv) consumption of
metabolic intermediates for biomass production based on measured
growth rate and biomass component contents. Specifically, [researchers]
measured the DNA, RNA, and protein contents to be equal to 11, 14,
and 88 µg/µl cells, respectively, in iBMK cells. Cellular demands for
acetyl-CoA based on steady-state fatty acid concentrations, cellular
growth rate, and fatty acid uptake rates are shown in Supplementary
Table 5 and (v) measured flux in the serine biosynthesis pathway." |