Document Detail


xCt cystine transporter expression in HEK293 cells: pharmacology and localization.
MedLine Citation:
PMID:  11302733     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
xCT, the core subunit of the system x(c)(-) high affinity cystine transporter, belongs to a superfamily of glycoprotein-associated amino acid transporters. Although xCT was shown to promote cystine transport in Xenopus oocytes, little work has been done with mammalian cells (Sato, H., Tamba, M., Ishii, T., and Bannai, S. J. Biol. Chem. 274, 11455-11458, 1999). Therefore, we have constructed mammalian expression vectors for murine xCT and its accessory subunit 4F2hc and transfected them into HEK293 cells. We report that this transporter binds cystine with high affinity (81 microM) and displays a pharmacological profile expected for system x(c)(-). Surprisingly, xCT transport activity in HEK293 cells is not dependent on the co-expression of the exogenous 4F2hc. Expression of GFP-tagged xCT indicated a highly clustered plasma membrane and intracellular distribution suggesting the presence of subcellular domains associated with combating oxidative stress. Our results indicate that HEK293 cells transfected with the xCT subunit would be a useful vehicle for future structure-function and pharmacology experiments involving system x(c)(-).
Authors:
A Y Shih; T H Murphy
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  282     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2001 Apr 
Date Detail:
Created Date:  2001-04-16     Completed Date:  2001-05-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1132-7     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
Affiliation:
Department of Psychiatry, University of British Columbia, 4N1-2255 Wesbrook Mall, Vancouver, British Columbia, V6T 1Z3, Canada. thmurphy@interchange.ubc.edu
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Transport System y+*
Animals
Antigens, CD / biosynthesis,  genetics
Antigens, CD98
Biological Transport / physiology
Carrier Proteins / biosynthesis*,  genetics
Cell Line
Cell Membrane / metabolism
Cystine / metabolism
Genetic Vectors / genetics,  metabolism
Glutamic Acid / pharmacokinetics
Green Fluorescent Proteins
Humans
Intracellular Fluid / metabolism
Kidney / cytology,  metabolism*
Luminescent Proteins / genetics
Mice
Oxidative Stress / physiology
Protein Subunits
Recombinant Fusion Proteins / biosynthesis,  genetics
Transfection
Chemical
Reg. No./Substance:
0/Amino Acid Transport System y+; 0/Antigens, CD; 0/Antigens, CD98; 0/Carrier Proteins; 0/Luminescent Proteins; 0/Protein Subunits; 0/Recombinant Fusion Proteins; 0/SLC7A11 protein, human; 0/Slc7a11 protein, mouse; 147336-22-9/Green Fluorescent Proteins; 56-86-0/Glutamic Acid; 56-89-3/Cystine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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