RidA proteins are conserved enamine/imine deaminases found in all domains of life. In both prokaryotic and eukaryotic model organisms, deletion of ridA causes a number of phenotypes, including growth defects. Accumulation of 2-aminoacrylate (2AA) occurs in the absence of RidA and results in the respective phenotypes. The reactive 2AA enamine attacks pyridoxal 5’-phosphate (PLP, a B6 vitamer) in the active site of PLP-dependent enzymes, and covalently inactivates the enzyme. The metabolic impact resulting from damage to PLP-dependent enzymes varies depending on the specific metabolic needs of the organism. The role of RidA in cellular metabolism has been best characterized in prokaryotes such as Salmonella enterica, in which its 2AA deaminase activity was first identified. Even among closely related prokaryotes, consequences of ridA lesions vary, from inconsequential to crippling. Disruptions caused by 2AA accumulation are dependent on the needs of the organism and metabolic flux through relevant pathways. The work described herein was undertaken to improve understanding of the RidA paradigm in the eukaryotic model organism Saccharomyces cerevisiae, and by extension eukaryotes in general. The S. cerevisiae genome encodes two RidA homologs (Mmf1, Hmf1), but only one generates known phenotypes when deleted. The mitochondrially localized Mmf1 protein was previously shown to prevent 2AA accumulation and deleterious effects. Work presented here built upon prior knowledge to identify Hem1, a PLP-dependent enzyme, as a definitive target of 2AA in the S. cerevisiae mitochondria. Damage to Hem1 accounts for a portion of the disruptive effects of 2AA accumulation, including decreasing intracellular heme levels. Beyond the study on Hem1, a role for the cytosolic RidA homolog, Hmf1, was investigated. In total this work not only furthers our understanding of cellular metabolism in yeast, but also lays the groundwork for defining the role of RidA homologs in complex organisms, about which little is known. Yeast bears many similarities to higher eukaryotes, not least of which is the presence of mitochondria and a mitochondrially localized RidA homolog. Having identified a previously unknown impact of 2AA accumulation on heme biosynthesis, we make the reasonable assumption that this may occur in the mitochondria of other eukaryotic organisms.