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Dopamine Transporter Interactome Reveals Differential Interaction with DJ-1 when Exposed to METH

Graduate #10
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Natasha Berryman - Fisk University
Co-Author(s): Shalonda M. Ingram, Tanu Rana, and J. Shawn Goodwin, Meharry Medical College



The dopamine (DA) transporter (DAT) is essential for the reuptake of released dopamine (DA) in the brain. DAT is one of the main targets for psychostimulants leading to a disruption of DA homeostasis. We hypothesized that DAT interacting proteins are altered in the presence of methamphetamine (METH) compared to control conditions leading to DAT dysregulation. We used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to examine the interacting proteome of DAT. DAT-expressing cells were treated for 30 minutes with 10 μM METH or mock treated for control. Subsequently, a DAT monoclonal antibody was used to immunoprecipitate DAT and pull down DAT associated proteins. Our LC/MS-MS data provides evidence that differential binding proteins are involved in DAT-mediated drug response. Interestingly, our study revealed that DJ-1 interacts with DAT in control conditions, but does not interact with DAT after treatment with METH. DJ-1, also known as PARK7, inhibits the aggregation of α-synuclein and interacts with DAT to increase DA reuptake.(1,2) Although further studies are needed examine the functional consequences of these findings, our data could point to a previously unknown mechanism by which METH alters DA homeostasis and leads to early onset of Parkinson’s Disease in METH abusers.(3)
1. Shendelman S, Jonason A, Martinat C, Leete T, Abeliovich A. DJ-1 is a redox-dependent molecular chaperone that inhibits alpha-synuclein aggregate formation. PLoS Biol. 2004 Nov; 2(11): e362.
2. Luk B, Mohammed M, Liu F, Lee FJ. A Physical Interaction between the Dopamine Transporter and DJ-1 Facilitates Increased Dopamine Reuptake. PLoS One. 2015; 10(8): e0136641
3. Curtin K, Fleckenstein AE, Robison RJ, Crookston MJ, Smith KR, Hanson GR. Methamphetamine/amphetamine abuse and risk of Parkinson’s disease in Utah: a population-based assessment. Drug Alcohol Depend. 2015 Jan 1;146:30-8.

Funder Acknowledgement(s): NIH grants U54 MD007593, G12 MD007586, U54 CA163069, R24 DA036420

Faculty Advisor: J. Shawn Goodwin, jgoodwin@mmc.edu

Role: Under the training and direction of Ingram, Rana and Goodwin: Used LC-MS/MS to examine the proteome of DAT; Prepared and treated DAT-expressing cells according to study design; Used DAT monoclonal antibody to immunoprecipitate DAT in DAT-expressing cells; Conducted pull-down DAT assays; Collected and analyzed data.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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