Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Kidney Int
2005 Aug 01;682:653-64. doi: 10.1111/j.1523-1755.2005.00443.x.
Show Gene links
Show Anatomy links
Cystine and glutamate transport in renal epithelial cells transfected with human system x(-) (c).
Bridges CC
,
Zalups RK
.
???displayArticle.abstract???
BACKGROUND: System x(-) (c) is a heterodimeric transporter, comprised of a light chain, xCT, and heavy chain, 4F2hc, which mediates the sodium-independent exchange of cystine and glutamate at the plasma membrane. In the current study we tested the hypothesis that stable transfection of Madin-Darby canine kidney (MDCK) cells with human xCT and 4F2hc results in the expression of functional system x(-) (c).
METHODS: MDCK cells were transfected stably with human clones for xCT and 4F2hc. Analyses of time- and temperature-dependence, saturation kinetics, and substrate specificity of l-cystine and l-glutamate transport were carried out in control and xCT-4F2hc-transfected MDCK cells. We also measured the uptake of l-cystine in Xenopus oocytes expressing human xCT and/or 4F2hc or xCT and/or rBAT (a heavy chain homologous to 4F2hc).
RESULTS: All of the different sets of data revealed that transport of l-cystine and l-glutamate increased significantly (twofold to threefold) in the MDCK cells subsequent to transfection with xCT-4F2hc. Moreover, uptake of l-cystine also increased (about tenfold) in Xenopus oocytes expressing hxCT and h4F2hc. Biochemical analyses of l-cystine uptake in oocytes verified our findings in the transfected MDCK cells. Interestingly, in oocytes injected with rBAT with or without xCT, uptake of l-cystine was significantly greater than that in water-injected oocytes.
CONCLUSION: Our findings indicate that stable transfection of MDCK cells with xCT and 4F2hc results in a cell-line expressing a functional system x(-) (c) transporter that can utilize l-cystine and l-glutamate as substrates. This study apparently represents the first stable transfection of a mammalian cell line with system x(-) (c).
???displayArticle.pubmedLink???
16014042
???displayArticle.pmcLink???PMC2409290 ???displayArticle.link???Kidney Int ???displayArticle.grants???[+]
Aslamkhan,
Human renal organic anion transporter 1-dependent uptake and toxicity of mercuric-thiol conjugates in Madin-Darby canine kidney cells.
2003, Pubmed,
Xenbase
Aslamkhan,
Human renal organic anion transporter 1-dependent uptake and toxicity of mercuric-thiol conjugates in Madin-Darby canine kidney cells.
2003,
Pubmed
,
Xenbase
Bannai,
Role of proton dissociation in the transport of cystine and glutamate in human diploid fibroblasts in culture.
1981,
Pubmed
Bannai,
Transport of cystine and cysteine and cell growth in cultured human diploid fibroblasts: effect of glutamate and homocysteate.
1982,
Pubmed
Bannai,
Transport of cystine and cysteine in mammalian cells.
1984,
Pubmed
Bannai,
Exchange of cystine and glutamate across plasma membrane of human fibroblasts.
1986,
Pubmed
Bannai,
Enhancement of glutathione levels in mouse peritoneal macrophages by sodium arsenite, cadmium chloride and glucose/glucose oxidase.
1991,
Pubmed
Bannai,
Transport interaction of L-cystine and L-glutamate in human diploid fibroblasts in culture.
1980,
Pubmed
Bassi,
Identification and characterisation of human xCT that co-expresses, with 4F2 heavy chain, the amino acid transport activity system xc-.
2001,
Pubmed
,
Xenbase
Bauch,
Apical heterodimeric cystine and cationic amino acid transporter expressed in MDCK cells.
2002,
Pubmed
,
Xenbase
Boerner,
Polarity of neutral amino acid transport and characterization of a broad specificity transport activity in a kidney epithelial cell line, MDCK.
1986,
Pubmed
Bridges,
Structure, function, and regulation of human cystine/glutamate transporter in retinal pigment epithelial cells.
2001,
Pubmed
Bridges,
Homocysteine, system b0,+ and the renal epithelial transport and toxicity of inorganic mercury.
2004,
Pubmed
Bridges,
Induction of cystine-glutamate transporter xc- by human immunodeficiency virus type 1 transactivator protein tat in retinal pigment epithelium.
2004,
Pubmed
Bridges,
Mercuric conjugates of cysteine are transported by the amino acid transporter system b(0,+): implications of molecular mimicry.
2004,
Pubmed
Cariappa,
Stable polarized expression of hCAT-1 in an epithelial cell line.
2002,
Pubmed
,
Xenbase
Chillarón,
Heteromeric amino acid transporters: biochemistry, genetics, and physiology.
2001,
Pubmed
Foreman,
Characteristics of cystine uptake by cultured LLC-PK1 cells.
1988,
Pubmed
Gukasyan,
Regulation of L-cystine transport and intracellular GSH level by a nitric oxide donor in primary cultured rabbit conjunctival epithelial cell layers.
2003,
Pubmed
Kim,
Human cystine/glutamate transporter: cDNA cloning and upregulation by oxidative stress in glioma cells.
2001,
Pubmed
,
Xenbase
Makowske,
Contrasts in transport systems for anionic amino acids in hepatocytes and a hepatoma cell line HTC.
1982,
Pubmed
Miura,
Cystine uptake and glutathione level in endothelial cells exposed to oxidative stress.
1992,
Pubmed
Ochi,
Arsenic compound-induced increases in glutathione levels in cultured Chinese hamster V79 cells and mechanisms associated with changes in gamma-glutamylcysteine synthetase activity, cystine uptake and utilization of cysteine.
1997,
Pubmed
Palacin,
The amino acid transport system b(o,+) and cystinuria.
2001,
Pubmed
Palacín,
Molecular biology of mammalian plasma membrane amino acid transporters.
1998,
Pubmed
Patel,
Differentiation of substrate and non-substrate inhibitors of transport system xc(-): an obligate exchanger of L-glutamate and L-cystine.
2004,
Pubmed
Rajan,
Cloning and expression of a b(0,+)-like amino acid transporter functioning as a heterodimer with 4F2hc instead of rBAT. A new candidate gene for cystinuria.
1999,
Pubmed
,
Xenbase
Sato,
Induction of cystine transport via system x-c and maintenance of intracellular glutathione levels in pancreatic acinar and islet cell lines.
1998,
Pubmed
Sato,
Transcriptional control of cystine/glutamate transporter gene by amino acid deprivation.
2004,
Pubmed
Sato,
Molecular cloning and expression of human xCT, the light chain of amino acid transport system xc-.
2000,
Pubmed
,
Xenbase
Sato,
Cloning and expression of a plasma membrane cystine/glutamate exchange transporter composed of two distinct proteins.
1999,
Pubmed
,
Xenbase
Takada,
Transport of cystine in isolated rat hepatocytes in primary culture.
1984,
Pubmed
Tomi,
Induction of xCT gene expression and L-cystine transport activity by diethyl maleate at the inner blood-retinal barrier.
2002,
Pubmed
Tomi,
Expression and regulation of L-cystine transporter, system xc-, in the newly developed rat retinal Müller cell line (TR-MUL).
2003,
Pubmed
Verrey,
Novel renal amino acid transporters.
2005,
Pubmed
Wang,
Expression of the activity of cystine/glutamate exchange transporter, system x(c)(-), by xCT and rBAT.
2003,
Pubmed
,
Xenbase
Warren,
The Lathyrus excitotoxin beta-N-oxalyl-L-alpha,beta-diaminopropionic acid is a substrate of the L-cystine/L-glutamate exchanger system xc-.
2004,
Pubmed
Watanabe,
Induction of cystine transport activity in mouse peritoneal macrophages.
1987,
Pubmed
