Abstract 1116 Characterizing enzymatic control of the enzyme mitoNEET

MitoNEET is a [2Fe-2S] cluster-containing redox active mitochondrial protein that was identified as a target for the development of drugs to treat type-2 diabetes, cancer, and other diseases. However, mitoNEET's unclear functional role in cells remains a roadblock for therapeutic design. Recent research from the Konkle Lab has revealed that the Lys55 residue of mitoNEET is particularly reactive because it is in a hydrogen-bonding network with the histidine that ligates the cluster. As such, it is selectively modified by the reactive electrophile 4-oxononenal and the enzymatic cofactor pyridoxal phosphate (PLP). Once loaded with PLP, mitoNEET catalyzes the transamination between cysteine and 2-oxoglutarate to produce 3-mercaptopyruvate and glutamate. Analysis of the metabolites was accomplished by derivatization followed by separation by either GC-MS or HPLC. We have further investigated the function of mitoNEET as a transaminase enzyme in the presence of various ligands already known to bind to the protein. This work further characterizes mitoNEET as a transaminase enzyme that contributes to the dynamic cellular thiolome. NSF funding CHE1806266,HRD-1618408,EES-2308500