Chemistry and Biochemistry
Bayer School of Natural and Environmental Sciences
John F. Stolz
Michael Van Stipdonk
Jesus Tejero Bravo
mARC, Mass spectrometry, Molybdenum enzymes, Protein-protein interactions
Newly discovered pterin-molybdoenzymes mammalian proteins, mARC1, and mARC2, are though to activate pro-drug, metabolize mutated base pairs, and producing nitric oxide from nitrite. However, the physiological function of the mARC proteins remain unclear. It is hypothesized that identifying partner proteins through protein-protein interactions can provide evidence to their physiological function. This study aimed to further investigate the potential function of mARC proteins through the identification of protein-protein interaction in human cell lysate, utilizing co-immunoprecipitation, crosslinking reagent, and pull-down assays. Several proteins (e.g. mitogen-activated protein kinase 5, phosphatidylinositol 4-phospahte 3-kinase C2 domain-containing subunit beta, ADP/ATP translocase 1, and myotubularin) were identified as interacting partners of mARC1, and mARC2. These interacting proteins are affiliated with various KEGG signaling pathways. From the interacting proteins we infer, mARC1 and mARC2 may be involved in the regulation of nitric oxide signaling under hypoxic. Reverse co-immunoprecipitation experiments using select proteins from the pathways detected mARC proteins. The investigation was then expanded to include pig liver mitochondrial fraction and rat liver cell lysate, in which all experimental conditions identified mARC proteins using reverse co-immunoprecipitation.
Thomas, J. (2016). Protein-Protein Interactions of Human Mitochondrial Amidoxime-Reducing Component in Mammalian Cells (Doctoral dissertation, Duquesne University). Retrieved from https://dsc.duq.edu/etd/61