JPET xPharm- The Comprehensive Pharmacology Reference

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Doolen, S.
Right arrow Articles by Zahniser, N. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Doolen, S.
Right arrow Articles by Zahniser, N. R.

Vol. 296, Issue 3, 931-938, March 2001

Protein Tyrosine Kinase Inhibitors Alter Human Dopamine Transporter Activity in Xenopus Oocytes

Suzanne Doolen and Nancy R. Zahniser

Department of Pharmacology and Neuroscience Program, University of Colorado Health Sciences Center, Denver, Colorado

The dopamine (DA) transporter (DAT) regulates dopaminergic synaptic transmission by controlling extracellular levels of DA. Thus, understanding signaling mechanisms that alter DAT function is critical for understanding dopaminergic neurotransmission. We have expressed the human DAT (hDAT) in Xenopus laevis oocytes to test the hypothesis that protein tyrosine kinases (PTKs) acutely regulate DAT function by altering cell surface expression of the transporter. Using a relatively high concentration of DA (10 µM), we found that several PTK inhibitors, namely, genistein, lavendustin A, and tyrphostin 25 (10 µM), decreased DA uptake velocity by 58, 41, and 30% of control, respectively. Furthermore, genistein potently inhibited DA uptake with a Ki = 68 nM. Kinetic analysis confirmed that genistein decreased the Vmax of the DAT, with no change in Km. The effects of PTK inhibition on hDAT-associated currents were also measured. All three PTK inhibitors attenuated substrate transport-associated currents to similar extents as DA uptake. In contrast, the potent Src inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) did not significantly inhibit either DA uptake or transport-associated currents. PTK inhibitors decreased hDAT-associated leak currents, however in a more variable manner than for uptake and transport-associated currents. Genistein also decreased cell surface binding of [3H]WIN 35,428 to hDAT by 48% of control. Together, these data provide several lines of evidence suggesting that PTK inhibition rapidly reduces hDAT activity via redistribution of the transporter away from the cell surface. Thus, PTKs likely represent another component of cellular signaling cascades that acutely regulate neurotransmitter transporters.

0022-3565/01/2963-0931$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2001 by The American Society for Pharmacology and Experimental Therapeutics


This article has been cited by other articles:


Home page
 Mol. Pharmacol.Home page
Y. Wei, J. M. Williams, C. Dipace, U. Sung, J. A. Javitch, A. Galli, and C. Saunders
Dopamine Transporter Activity Mediates Amphetamine-Induced Inhibition of Akt through a Ca2+/Calmodulin-Dependent Kinase II-Dependent Mechanism
Mol. Pharmacol., March 1, 2007; 71(3): 835 - 842.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
B. G. Garcia, Y. Wei, J. A. Moron, R. Z. Lin, J. A. Javitch, and A. Galli
Akt Is Essential for Insulin Modulation of Amphetamine-Induced Human Dopamine Transporter Cell-Surface Redistribution
Mol. Pharmacol., July 1, 2005; 68(1): 102 - 109.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. M. Kahlig, J. A. Javitch, and A. Galli
Amphetamine Regulation of Dopamine Transport: COMBINED MEASUREMENTS OF TRANSPORTER CURRENTS AND TRANSPORTER IMAGING SUPPORT THE ENDOCYTOSIS OF AN ACTIVE CARRIER
J. Biol. Chem., March 5, 2004; 279(10): 8966 - 8975.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Sorkina, S. Doolen, E. Galperin, N. R. Zahniser, and A. Sorkin
Oligomerization of Dopamine Transporters Visualized in Living Cells by Fluorescence Resonance Energy Transfer Microscopy
J. Biol. Chem., July 18, 2003; 278(30): 28274 - 28283.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Z. Lin, P.-W. Zhang, X. Zhu, J.-M. Melgari, R. Huff, R. L. Spieldoch, and G. R. Uhl
Phosphatidylinositol 3-Kinase, Protein Kinase C, and MEK1/2 Kinase Regulation of Dopamine Transporters (DAT) Require N-terminal DAT Phosphoacceptor Sites
J. Biol. Chem., May 23, 2003; 278(22): 20162 - 20170.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
X.-L. Tang, E. Kodani, H. Takano, M. Hill, K. Shinmura, T. M. Vondriska, P. Ping, and R. Bolli
Protein tyrosine kinase signaling is necessary for NO donor-induced late preconditioning against myocardial stunning
Am J Physiol Heart Circ Physiol, April 1, 2003; 284(4): H1441 - H1448.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Maiya, K. J. Buck, R. A. Harris, and R. D. Mayfield
Ethanol-sensitive Sites on the Human Dopamine Transporter
J. Biol. Chem., August 16, 2002; 277(34): 30724 - 30729.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
A. M. Carneiro, S. L. Ingram, J.-M. Beaulieu, A. Sweeney, S. G. Amara, S. M. Thomas, M. G. Caron, and G. E. Torres
The Multiple LIM Domain-Containing Adaptor Protein Hic-5 Synaptically Colocalizes and Interacts with the Dopamine Transporter
J. Neurosci., August 15, 2002; 22(16): 7045 - 7054.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. D. Foster, B. Pananusorn, and R. A. Vaughan
Dopamine Transporters Are Phosphorylated on N-terminal Serines in Rat Striatum
J. Biol. Chem., July 5, 2002; 277(28): 25178 - 25186.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
S. Apparsundaram, U. Sung, R. D. Price, and R. D. Blakely
Trafficking-Dependent and -Independent Pathways of Neurotransmitter Transporter Regulation Differentially Involving p38 Mitogen-Activated Protein Kinase Revealed in Studies of Insulin Modulation of Norepinephrine Transport in SK-N-SH Cells
J. Pharmacol. Exp. Ther., November 1, 2001; 299(2): 666 - 677.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 2001 by the American Society for Pharmacology and Experimental Therapeutics.