Evidence map›Paper›PMID 24753048›Full record

ArticleMolecular pharmacology2014

An N-terminal threonine mutation produces an efflux-favorable, sodium-primed conformation of the human dopamine transporter.

Rheaclare Fraser, Yongyue Chen, Bipasha Guptaroy, Kathryn D Luderman, Stephanie L Stokes, Asim Beg, Louis J DeFelice, Margaret E Gnegy

Open access · bronzeAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Molecular pharmacology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.

0numbers the graph read from it
0cells of the map it votes in
7citing papers in PubMed
0.7field-weighted citation impact, top 29% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

7 citing papers in PubMed, 11 citations in OpenAlex.

  1. Article
  2. Review
  3. Review
  4. Chloride requirement for monoamine transporters.Pflugers Archiv : European journal of physiology · 2016
    Review
  5. Dopamine Transporter Activity Is Modulated by α-Synuclein.The Journal of biological chemistry · 2015
    Article
  6. Article
  7. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 1 country.

Rheaclare FraserDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Yongyue ChenDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Bipasha GuptaroyDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Kathryn D LudermanDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Stephanie L StokesDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Asim BegDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Louis J DeFeliceDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia.
Margaret E GnegyDepartment of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.F., B.G., K.D.L., S.L.S., A.B., M.E.G.); and Departments of Psychiatry (Y.C.) and Physiology and Biophysics (L.J.D.), Virginia Commonwealth University, Richmond, Virginia pgnegy@umich.edu.
University of Michigan–Ann Arbor · USVirginia Commonwealth University · US

Funding

INTERDEPARTMENTAL TRAINING IN PHARMACOLOGICAL SCIENCEST32GM007767 · NIGMS · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI ISOM, LORI L. · 1985 to 2019
$10.2M
NIDA Training Program in Neuroscience-Administrative SupplementT32DA007281 · NIDA · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI Shelly Beth Flagel · 1995 to 2026
$5.7M
PHARMACOLOGY OF DOPAMINE RELEASE BY AMPHETAMINER01DA011697 · NIDA · UNIVERSITY OF MICHIGAN AT ANN ARBOR · PI GNEGY, MARGARET E · 2000 to 2016
$5.3M
Methamphetamine regulates the dopamine transporter via an intracellular mechanismR01DA026947 · NIDA · UNIVERSITY OF FLORIDA · PI KHOSHBOUEI, HABIBEH · 2010 to 2019
$3.5M
Synthetic cathinones: a new class of illicit drugs affecting DAT & SERTR01DA033930 · NIDA · VIRGINIA COMMONWEALTH UNIVERSITY · PI ELTIT, JOSE MIGUEL, GLENNON, RICHARD A · 2012 to 2016
$2.7M
Amphetamine Acts as a Molecular Stent in the Dopamine TransporterRC1DA028112 · NIDA · VIRGINIA COMMONWEALTH UNIVERSITY · PI DE FELICE, LOUIS J · 2009 to 2010
$1.0M
NIDA NIH HHS 5R01-DA011697NIDA NIH HHS 5R01-DA026947NIDA NIH HHS 5RC1-DA028112NIDA NIH HHS 5T32-DA007281NIDA NIH HHS R01 DA011697NIDA NIH HHS R01 DA026947NIDA NIH HHS R01 DA033930NIDA NIH HHS RC1 DA028112NIDA NIH HHS T32 DA007281NIGMS NIH HHS T32 GM007767
6 · The paper itself

Abstract

The dopamine transporter (DAT) reversibly transports dopamine (DA) through a series of conformational transitions. Alanine (T62A) or aspartate (T62D) mutagenesis of Thr62 revealed T62D-human (h)DAT partitions in a predominately efflux-preferring conformation. Compared with wild-type (WT), T62D-hDAT exhibits reduced [(3)H]DA uptake and enhanced baseline DA efflux, whereas T62A-hDAT and WT-hDAT function in an influx-preferring conformation. We now interrogate the basis of the mutants' altered function with respect to membrane conductance and Na(+) sensitivity. The hDAT constructs were expressed in Xenopus oocytes to investigate if heightened membrane potential would explain the efflux characteristics of T62D-hDAT. In the absence of substrate, all constructs displayed identical resting membrane potentials. Substrate-induced inward currents were present in oocytes expressing WT- and T62A-hDAT but not T62D-hDAT, suggesting equal bidirectional ion flow through T62D-hDAT. Utilization of the fluorescent DAT substrate ASP(+) [4-(4-(dimethylamino)styryl)-N-methylpyridinium] revealed that T62D-hDAT accumulates substrate in human embryonic kidney (HEK)-293 cells when the substrate is not subject to efflux. Extracellular sodium (Na(+) e) replacement was used to evaluate sodium gradient requirements for DAT transport functions. The EC50 for Na(+) e stimulation of [(3)H]DA uptake was identical in all constructs expressed in HEK-293 cells. As expected, decreasing [Na(+)]e stimulated [(3)H]DA efflux in WT- and T62A-hDAT cells. Conversely, the elevated [(3)H]DA efflux in T62D-hDAT cells was independent of Na(+) e and commensurate with [(3)H]DA efflux attained in WT-hDAT cells, either by removal of Na(+) e or by application of amphetamine. We conclude that T62D-hDAT represents an efflux-willing, Na(+)-primed orientation-possibly representing an experimental model of the conformational impact of amphetamine exposure to hDAT.

Indexed as

AnimalsBiological TransportCell LineDopamine Plasma Membrane Transport ProteinsHEK293 CellsHumansMembrane GlycoproteinsMembrane PotentialsMembrane Transport ProteinsMutationOocytesSodiumThreonineXenopusDopamine Plasma Membrane Transport ProteinsMembrane GlycoproteinsMembrane Transport ProteinsSLC6A3 protein, humanSodiumThreonine

Identifiers

PMID24753048
PMCPMC4053999
OpenAlexW2169404124

What OpenQuestion holds

Textmetadata
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.