Reserve Flux Capacity in the Pentose Phosphate Pathway by NADPH Binding Is Conserved across Kingdoms

All organisms evolved defense mechanisms to counteract oxidative stress and buildup of reactive oxygen species (ROS). To test whether a potentially conserved mechanism exists for the rapid response, we investigated immediate metabolic dynamics of Escherichia coli, yeast, and human dermal fibroblasts to oxidative stress that we found to be conserved between species. To elucidate the regulatory mechanisms that implement this metabolic response, we developed mechanistic kinetic models for each organism's central metabolism and systematically tested activation and inactivation of each irreversible reaction by each metabolite. This ensemble modeling predicts in vivo relevant metabolite-enzyme interactions based on their ability to quantitatively describe metabolite dynamics. All three species appear to inhibit their oxidative pentose phosphate pathway during normal growth by the redox cofactor NADPH and relieve this inhibition to increase the pathway flux for detoxification of ROS during stress, with the sole exception of yeast when exposed to high levels of stress. [Display omitted] •Characterization of immediate metabolic response to oxidative stress•The metabolic response in glycolysis and PP pathway depends on stress severity•Identification of NADPH feedback inhibition on G6PDH as key regulatory interaction•The identified oxidative stress regulatory interaction is conserved across kingdoms Biological Sciences; Metabolism; Systems Biology; Metabolic Flux Analysis; Metabolomics; Computational Biology; Bioinformatics