Metabolism, a fundamental aspect of all biological systems, encompasses the intricate network of chemical reactions that occur within a living organism to maintain life. Metabolism interconnects a broad range of crucial biological processes including energy production, cellular repair, growth, and response to environmental changes. The study of metabolism not only provides insights into basic biological functions but also aids in understanding and treating metabolic disorders and diseases like cancer, diabetes, aging, and obesity. As a reflection of the diverse aspects of metabolism, this thesis outlines a variety of studies centered around regulatory signaling pathways that arise from reactive metabolites of the central, highly conserved glycolytic pathway to understand both fundamental aspects of cellular metabolism and disease mechanisms. It delves into the dynamics of 3-phosphoglyceryl-lysine (pgK) modifications in cyanobacteria, highlighting their oscillation in response to environmental light/dark cycles and impact on glycolytic enzymes and central carbon metabolism flux. It confirms the role of DJ-1 in detoxifying methylglyoxal as a glyoxalase, to set the foundation for future pathophysiological studies of the metabolism of neurodegenerative disorders. Furthermore, it explores the potential of histone deacetylases, specifically HDAC3, in regulating pgK modifications, and suggests potential research directions to study the broader functional implications of this mechanism. Lastly, this thesis maps how cancer cells respond to glycolytic perturbations, focusing in particular on the relationship between different forms of central glycolytic perturbation and the resulting stress responses; highlighting especially the role of central glycolysis as a nexus for redox stress signaling. Overall, this thesis aims to present findings that offer new insights into metabolic regulation and disease pathology by reactive metabolite post-translational modifications, with the goal of improving insight for next generation therapeutic approaches against metabolically aberrant diseases.