Dopamine transporter phosphorylation site threonine 53 regulates Research Paper

Dopamine transporter phosphorylation site threonine 53 regulates substrate reuptake and amphetamine-stimulated efflux - Research Paper Example (29702). Activities that lead to the reuptake of DAT are crucial for the functionality of the dopaminergic neurons. Dopaminergic disorders, for instance, schizophrenia and depression among others may come about as a result of dysregulation of the transport of DAT, which causes an inequality of dopamine. Commonly abused substances including drugs such as cocaine and amphetamine are often targeted by DAT. In addition, therapeutic agents that treat dopamine complications are also targeted by DAT. Specifically, AMPH and its congeners induce multiple short-term and long-term effects on DAT. The mechanism behind the efflux is related with transporter-generated currents. Various techniques have been used to determine the precise phosphorylation site of DAT. It has been established that that a recombinant peptide containing N-terminal residues 1–65 of rDAT underwent phosphorylation outside living tissues in the presence of proline-guided kinases. It was also established that the precise phosphorylation site in heterologously expressed protein was a threonine residue, which was located at position 53. However, it was vital to ascertain the phosphorylation site using other techniques. The researchers in this paper aimed at determining the role of dopamine transporter phosphorylation site threonine 53 on the reabsorption of the substrate and amphetamine-triggered efflux Foster et al. (29702). The researchers make use of mass spectrometry and a novel antibody that is specific to phosphates to ascertain the presence of DAT phosphorylation at Thr53 in striatal tissue obtained from rodents as well as heterologous expression systems. Cell culture and dopamine transporter mutagenesis was done using Lewis carcinoma cells that were stably expressing WTrDAT (rDAT-LLCPK) (31) or T53A or T53D rDAT and were maintained in alpha minimum essential medium. Dulbeccos modified