Premium
Dynamic Interaction of α‐synuclein and dopamine transporter at the plasma membrane
Author(s) -
Butler Brittany Renee,
Goodwin Shawn,
Khoshbouei Habibeh
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.844.5
Subject(s) - dopamine transporter , immunocytochemistry , fluorescence recovery after photobleaching , microbiology and biotechnology , dopamine plasma membrane transport proteins , biophysics , chemistry , alpha synuclein , membrane , transporter , cell membrane , dopamine , membrane protein , biology , neuroscience , biochemistry , parkinson's disease , endocrinology , medicine , gene , disease
The dopamine transporter (DAT) regulates the dimension and duration of DAergic transmission in the brain. We and others have shown that overexpression of α‐synuclein alters DAT activity. Previous studies have shown that α‐synuclein interacts at the C‐terminus of DAT, but the functional consequences of this interaction are not well known. This study examines the hypotheses that α‐synuclein interacts with DAT at the plasma membrane and this interaction alters DAT trafficking. To address these hypotheses we used Immunocytochemistry, live cell imaging, and co‐patching. The immunocytochemistry experiments in CHO cells co‐expressing YFP‐DAT and α‐synuclein suggest these proteins co‐localize at the plasma membrane. We found that although α‐synuclein is distributed homogenously in cells not expressing DAT, the intensity profile of α‐synuclein peaks at the plasma membrane of DAT cells, overlapping with that of DAT at the plasma membrane. Co‐patching experiments in living cells expressing YFP‐HA‐DAT +/− CFP‐α‐synuclein confirm co‐localization of the proteins at the membrane. We found a dynamic interaction between DAT and α‐synuclein and the recruitment of α‐synuclein to the plasma membrane. Fluorescence Recovery After Photobleaching experiments show that DAT decreases the lateral mobility of α‐synuclein. These results suggest the interaction between DAT and α‐synuclein is transient and dynamic.