Glimpse into the Dimerization, Distribution and Interaction Dynamics of ATP7A using Super Resolution Imaging
MetadataShow full item record
Copper is a very important component in the body for various cellular functions but an imbalance in its levels can be very toxic to the body. ATP7A and ATP7B are important to provide instructions in making proteins that regulate copper levels in the body. Despite ATP7A and ATP7B showing high homology they perform different functions that are not fully understood yet. Recently, ATP7B has been reported as a dimer in the cells, suggesting dimerization as a potential regulatory mechanism. Therefore, this research focuses on testing if the ATP7A can dimerize similar to the ATP7B and also aim to understand the sub-cellular distribution as well as the interaction dynamics of the dimer complex. To achieve that, we want to use signal molecule super-resolution imaging technique to visualize the dimerization and spatial temporal behaviors of ATP7A under different Cu conditions. For this purpose, I generated fluorescent protein (FP) tagged ATP7A constructs for mammalian cell expression. After transfecting the construct into COS7 cells, the expression and Cu responsibility of FP tagged ATP7A was characterized by Western blotting and fluorescent microscopy imaging respectively in fixed cells. With successful FP-tagging on ATP7A, we are ready for further single molecule imaging in live cells.