Dopamine transporter site-directed mutations differentially alter substrate transport and cocaine binding.

  title={Dopamine transporter site-directed mutations differentially alter substrate transport and cocaine binding.},
  author={S. Kitayama and S. Shimada and H. Xu and L. Markham and D. Donovan and G. Uhl},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  pages={7782 - 7785}
  • S. Kitayama, S. Shimada, +3 authors G. Uhl
  • Published 1992
  • Medicine, Biology
  • Proceedings of the National Academy of Sciences of the United States of America
Polar amino acids lying within three hydrophobic regions of the dopamine transporter (DAT) are analogous to those important for ligand recognition by catecholamine receptors. Possible functional significance of these amino acids was examined by expressing DAT cDNAs mutated in these polar residues. Replacement of aspartate at position 79 with alanine, glycine, or glutamate dramatically reduced uptake of [3H]dopamine and the tritium-labeled Parkinsonism-inducing neurotoxin 1-methyl-4… Expand
Dopamine transporter: transmembrane phenylalanine mutations can selectively influence dopamine uptake and cocaine analog recognition.
Phenylalanine residues lying in putative DAT transmembrane (TM) domains were hypothesized to contribute to aromatic and/or cationic interactions among DAT, dopamine, and cocaine. Expand
Dopamine uptake and cocaine binding mechanisms: the involvement of charged amino acids from the transmembrane domains of the human dopamine transporter.
The wild type human dopamine transporter (DAT) and five DAT mutants were transfected into COS-7 cells and their ability to uptake dopamine or to bind cocaine was examine three days later, and all mutants displayed a reduction or complete loss of the maximal velocity of dopamine transport. Expand
Dopamine transporter tryptophan mutants highlight candidate dopamine- and cocaine-selective domains.
These data, current provisional DAT structural models, and results from parallel studies of other mutants identify candidate dopamine-selective DAT domains for transmembrane dopamine permeation and regions in which mutations selectively lower CFT affinities. Expand
Cocaine affinity decreased by mutations of aromatic residue phenylalanine 105 in the transmembrane domain 2 of dopamine transporter.
The functional cocaine-insensitive mutants provide tools for the study of the mechanism of cocaine addiction and suggest that F105 in mDAT contributes to high-affinity cocaine binding. Expand
Tyrosine-533 of rat dopamine transporter: involvement in interactions with 1-methyl-4-phenylpyridinium and cocaine.
Tyrosine-533 may be important for the DAT function and for species differences in transporter functions, including differential sensitivities to cocaine and 1-methyl-1,2,3,6-tetrahydropyridine in humans and rats. Expand
Dissociation of high-affinity cocaine analog binding and dopamine uptake inhibition at the dopamine transporter.
A complex role for D79 is revealed in determining substrate specificity and high-affinity binding of DAT inhibitors and it is proposed that at least two discrete inhibitor-binding DAT conformations or populations exist and that the DAT conformation/population responsible for inhibitor high-Affinity binding is less responsible for dopamine uptake. Expand
Importance of valine at position 152 for the substrate transport and 2beta-carbomethoxy-3beta-(4-fluorophenyl)tropane binding of dopamine transporter.
Replacement of isoleucine, residue 152 of chimera hb3 (bovine DAT sequence), with valine, the human DAT residue at the identical position, remarkably restored the substrate transport and CFT binding to 76% to 98% of thehuman DAT values. Expand
The Third Transmembrane Domain of the Serotonin Transporter Contains Residues Associated with Substrate and Cocaine Binding*
The results suggest that Ile-172, Tyr-176 and Il-179 are on one face of an α-helical transmembrane element, and that I Le-172 and Tyr- 176 are in proximity to the binding site for 5-HT and cocaine. Expand
Recognition of Benztropine by the Dopamine Transporter (DAT) Differs from That of the Classical Dopamine Uptake Inhibitors Cocaine, Methylphenidate, and Mazindol as a Function of a DAT Transmembrane 1 Aspartic Acid Residue
Distinguishing the precise mechanism of action of this DAT inhibitor with relatively low abuse liability from that of cocaine may be attainable using DAT mutagenesis and other structure-function studies, opening the door to rational design of therapeutic agents for cocaine abuse. Expand
Dopamine transporter proline mutations influence dopamine uptake, cocaine analog recognition, and expression
  • Z. Lin, M. Itokawa, G. Uhl
  • Chemistry, Medicine
  • FASEB journal : official publication of the Federation of American Societies for Experimental Biology
  • 2000
This work reports production of mutants substituting alanine and/or glycine residues for 16 prolines located in or near putative DAT TM domains that identify DAT regions likely for dopamine translocation and for recognition of dopamine and cocaine. Expand