 |
||||
|
||||
|
||||
|
||||
The Journal of Neuroscience, June 1, 2003, 23(11):4470-4478
Previous Article | Next Article
A Mutation in the Human Norepinephrine Transporter Gene (SLC6A2) Associated with Orthostatic Intolerance Disrupts Surface Expression of Mutant and Wild-Type Transporters
Maureen K. Hahn,1,2 David Robertson,2,3,4 andRandy D. Blakely1,2,4 1 Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8548, 2 Center for Molecular Neuroscience, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8548, 3 Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8548, and 4 Autonomic Dysfunction Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8548
The norepinephrine transporter (NET) mediates reuptake of norepinephrine released from neurons, and, as such, it is an important regulator of noradrenergic neurotransmission. Recently, our laboratory reported a polymorphism in the human NET (hNET) gene A457P in an individual with the autonomic disorder orthostatic intolerance (OI). The presence of the hNET-A457P allele tracked with elevated heart rates and plasma NE levels in family members. hNET-A457P lacks >98% transport activity in several heterologous expression systems. In the present work, Western blot and biotinylation analyses performed in transiently transfected COS-7 cells revealed impairment in processing of hNET-A457P to the fully glycosylated form and a decrease in surface expression to
30% of hNET-wild type (hNET-wt). Because the hNET-A457P mutation is carried on a single allele in OI subjects, we examined the influence of cotransfection of hNET-wt and hNET-A457P and found that hNET-A457P exerts a dominant-negative effect on hNET-wt uptake activity. Experiments to determine oligomerization as a potential mechanism of the dominant-negative effect demonstrated that hNET-A457P coimmunoprecipitates with, and diminishes surface expression of, hNET-wt. These results reveal that hNET-A457P causes a conformational disruption that interferes with transporter biosynthetic progression and trafficking of both the mutant transporter and hNET-wt. These results elucidate a molecular mechanism for the disrupted NE homeostasis and cardiovascular function evident in OI patients with the hNET-A457P mutation.
Key words: norepinephrine; transporter; SLC6A2; orthostatic intolerance; trafficking; antidepressant
Received Sep. 6, 2002; revised Mar. 13, 2003; accepted Mar. 17, 2003.
This article has been cited by other articles:
I. Bartholomaus, L. Milan-Lobo, A. Nicke, S. Dutertre, H. Hastrup, A. Jha, U. Gether, H. H. Sitte, H. Betz, and V. Eulenburg
Glycine Transporter Dimers: EVIDENCE FOR OCCURRENCE IN THE PLASMA MEMBRANE
J. Biol. Chem., April 18, 2008; 283(16): 10978 - 10991.
[Abstract] [Full Text] [PDF]
J. N. Mason, D. C. Deecher, R. L. Richmond, G. Stack, P. E. Mahaney, E. Trybulski, R. C. Winneker, and R. D. Blakely
Desvenlafaxine Succinate Identifies Novel Antagonist Binding Determinants in the Human Norepinephrine Transporter
J. Pharmacol. Exp. Ther., November 1, 2007; 323(2): 720 - 729.
[Abstract] [Full Text] [PDF]
H. Farhan, V. Reiterer, V. M. Korkhov, J. A. Schmid, M. Freissmuth, and H. H. Sitte
Concentrative Export from the Endoplasmic Reticulum of the {gamma}-Aminobutyric Acid Transporter 1 Requires Binding to SEC24D
J. Biol. Chem., March 9, 2007; 282(10): 7679 - 7689.
[Abstract] [Full Text] [PDF]
P. Catarecha, M. D. Segura, J. M. Franco-Zorrilla, B. Garcia-Ponce, M. Lanza, R. Solano, J. Paz-Ares, and A. Leyva
A Mutant of the Arabidopsis Phosphate Transporter PHT1;1 Displays Enhanced Arsenic Accumulation
PLANT CELL, March 1, 2007; 19(3): 1123 - 1133.
[Abstract] [Full Text] [PDF]
T. Beuming, L. Shi, J. A. Javitch, and H. Weinstein
A Comprehensive Structure-Based Alignment of Prokaryotic and Eukaryotic Neurotransmitter/Na+ Symporters (NSS) Aids in the Use of the LeuT Structure to Probe NSS Structure and Function
Mol. Pharmacol., November 1, 2006; 70(5): 1630 - 1642.
[Abstract] [Full Text] [PDF]
V. M. Korkhov, M. Holy, M. Freissmuth, and H. H. Sitte
The Conserved Glutamate (Glu136) in Transmembrane Domain 2 of the Serotonin Transporter Is Required for the Conformational Switch in the Transport Cycle
J. Biol. Chem., May 12, 2006; 281(19): 13439 - 13448.
[Abstract] [Full Text] [PDF]
K.-H. Lee, B.-H. Ko, J.-Y. Paik, K.-H. Jung, J.-S. Bae, J.-Y. Choi, Y. S. Choe, and B.-T. Kim
Characteristics and Regulation of 123I-MIBG Transport in Cultured Pulmonary Endothelial Cells
J. Nucl. Med., March 1, 2006; 47(3): 437 - 442.
[Abstract] [Full Text] [PDF]
L. K. Henry, J. R. Field, E. M. Adkins, M. L. Parnas, R. A. Vaughan, M.-F. Zou, A. H. Newman, and R. D. Blakely
Tyr-95 and Ile-172 in Transmembrane Segments 1 and 3 of Human Serotonin Transporters Interact to Establish High Affinity Recognition of Antidepressants
J. Biol. Chem., January 27, 2006; 281(4): 2012 - 2023.
[Abstract] [Full Text] [PDF]
M. K. Hahn, M. S. Mazei-Robison, and R. D. Blakely
Single Nucleotide Polymorphisms in the Human Norepinephrine Transporter Gene Affect Expression, Trafficking, Antidepressant Interaction, and Protein Kinase C Regulation
Mol. Pharmacol., August 1, 2005; 68(2): 457 - 466.
[Abstract] [Full Text] [PDF]
A. E. Ruoho
How the Monoamine Transporter Garden Grows
Mol. Pharmacol., August 1, 2005; 68(2): 272 - 274.
[Abstract] [Full Text] [PDF]
J. W. Schwartz, G. Novarino, D. W. Piston, and L. J. DeFelice
Substrate Binding Stoichiometry and Kinetics of the Norepinephrine Transporter
J. Biol. Chem., May 13, 2005; 280(19): 19177 - 19184.
[Abstract] [Full Text] [PDF]
D. S. Goldstein, B. Eldadah, C. Holmes, S. Pechnik, J. Moak, and Y. Sharabi
Neurocirculatory Abnormalities in Chronic Orthostatic Intolerance
Circulation, February 22, 2005; 111(7): 839 - 845.
[Abstract] [Full Text] [PDF]
S. Seidel, E. A. Singer, H. Just, H. Farhan, P. Scholze, O. Kudlacek, M. Holy, K. Koppatz, P. Krivanek, M. Freissmuth, et al.
Amphetamines Take Two to Tango: an Oligomer-Based Counter-Transport Model of Neurotransmitter Transport Explores the Amphetamine Action
Mol. Pharmacol., January 1, 2005; 67(1): 140 - 151.
[Abstract] [Full Text] [PDF]
V. M. Korkhov, H. Farhan, M. Freissmuth, and H. H. Sitte
Oligomerization of the {gamma}-Aminobutyric Acid Transporter-1 Is Driven by an Interplay of Polar and Hydrophobic Interactions in Transmembrane Helix II
J. Biol. Chem., December 31, 2004; 279(53): 55728 - 55736.
[Abstract] [Full Text] [PDF]
N. R. Keller, A. Diedrich, M. Appalsamy, S. Tuntrakool, S. Lonce, C. Finney, M. G. Caron, and D. Robertson
Norepinephrine Transporter-Deficient Mice Exhibit Excessive Tachycardia and Elevated Blood Pressure With Wakefulness and Activity
Circulation, September 7, 2004; 110(10): 1191 - 1196.
[Abstract] [Full Text] [PDF]
M. J. Gallagher, L. Song, F. Arain, and R. L. Macdonald
The Juvenile Myoclonic Epilepsy GABAA Receptor {alpha}1 Subunit Mutation A322D Produces Asymmetrical, Subunit Position-Dependent Reduction of Heterozygous Receptor Currents and {alpha}1 Subunit Protein Expression
J. Neurosci., June 16, 2004; 24(24): 5570 - 5578.
[Abstract] [Full Text] [PDF]
L. D. Jayanthi, D. J. Samuvel, and S. Ramamoorthy
Regulated Internalization and Phosphorylation of the Native Norepinephrine Transporter in Response to Phorbol Esters: EVIDENCE FOR LOCALIZATION IN LIPID RAFTS AND LIPID RAFT-MEDIATED INTERNALIZATION
J. Biol. Chem., April 30, 2004; 279(18): 19315 - 19326.
[Abstract] [Full Text] [PDF]
D. L. Murphy, A. Lerner, G. Rudnick, and K.-P. Lesch
Serotonin Transporter: Gene, Genetic Disorders, and Pharmacogenetics
Mol. Interv., April 1, 2004; 4(2): 109 - 123.
[Abstract] [Full Text] [PDF]
|
|
|
|
 |
|