146 Chapter 5 Figure 3. The effects of amino-acid restriction on the peroxisomal β-oxidation. (A) Overall pathway flux in μmol · min ·mg of peroxisomal protein both in control and amino-acid restriction and amino-acid restriction supplemented with 100 μM DHA restricted conditions. (B) Enoyl-CoA and (C) Ketoacyl-CoA profiles in control and amino-acid restriction and amino-acid restriction supplemented with 100 μM DHA. Data represents simulations for 3 biological replicates (* indicates comparisons between control and -aa, # indicates comparisons between -aa and -aa + DHA, no indication means not significant) (*P < 0.05, **P < 0.01, #P<0.05 paired two-tailed t-test). We also simulated the effect of supplementing the amino-acid restricted organoids with docosahexaenoic acid (DHA). As we previously reported (Chapter 4), DHA supplementation to amino-acid restricted organoids prevented the loss of multiple peroxisomal and mitochondrial proteins in a dose-dependent manner, while it did not prevent the accumulation of TGs. When supplementing DHA, the flux of the pathway was significantly increased when compared to that of the amino-acid restricted organoids, and reached levels similar to those of the controls (Figure 3d). Moreover, the decrease in all enoyl and ketoacyl-CoA species was also prevented (Figure 3e and f), while no changes in the values of acyl-CoAs or acyl-carnitines were observed (Supplementary Figure 1). CONCLUSIONS AND OUTLOOK In this paper we presented a review on different enzymes involved in the peroxisomal β-oxidation of straight-chain fatty acids and used the data to generate (to the best of our knowledge) the first computational kinetic model
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