scholarly journals Overexpression of LAT1/CD98 Light Chain Is Sufficient to Increase System l-Amino Acid Transport Activity in Mouse Hepatocytes but Not Fibroblasts

2001 ◽  
Vol 276 (20) ◽  
pp. 16877-16884 ◽  
Author(s):  
William A. Campbell ◽  
Nancy L. Thompson
Keyword(s):  
Amino Acid ◽  
Light Chain ◽  
Amino Acid Transport ◽  
Acid Transport ◽  
Transport Activity ◽  
System L ◽  
Mouse Hepatocytes

scholarly journals A rapid method for the functional reconstitution of amino acid transport systems from rat liver plasma membranes. Partial purification of System A

1988 ◽  
Vol 255 (3) ◽  
pp. 963-969 ◽  
Author(s):  
A R Quesada ◽  
J D McGivan
Keyword(s):  
Amino Acid ◽  
Rapid Method ◽  
Amino Acid Transport ◽  
Plasma Membranes ◽  
Transport Systems ◽  
Partial Purification ◽  
Acid Transport ◽  
Transport Activity ◽  
System A ◽  
System L

A rapid method for the functional reconstruction of amino acid transport from liver plasma-membrane vesicles using the neutral detergent decanoyl-N-glucamide (‘MEGA-10’) is described. The method is a modification of that previously employed in this laboratory for reconstitution of amino acid transport systems from kidney brush-border membranes [Lynch & McGivan (1987) Biochem. J. 244, 503-508]. The transport activities termed ‘System A’, ‘System N’, and ‘System L’ are all reconstituted. The reconstitution procedure is rapid and efficient and is suitable as an assay for transport activity in studies involving membrane fractionation. By using this reconstitution procedure, System A transport activity was partially purified by lectin-affinity chromatography.

scholarly journals Expression of the surface antigen 4F2hc affects system-L-like neutral-amino-acid-transport activity in mammalian cells

1997 ◽  
Vol 324 (2) ◽  
pp. 535-541 ◽  
Author(s):  
Stefan BRÖER ◽  
Angelika BRÖER ◽  
Bernd HAMPRECHT
Keyword(s):  
Amino Acid ◽  
Surface Antigen ◽  
Amino Acid Transport ◽  
Cho Cells ◽  
Mammalian Cells ◽  
Primary Cultures ◽  
Transport Systems ◽  
Acid Transport ◽  
Transport Activity ◽  
System L

Mammalian cells possess a variety of amino acid-transport systems with overlapping substrate specificity. System L is one of the major amino acid-transport systems of non-epithelial cells. By expression cloning we have recently demonstrated that the surface antigen 4F2hc (CD98) is a necessary component for expression of system-L-like amino acid-transport activity in C6-BU-1 rat glioma cells [Bröer, Bröer and Hamprecht (1995) Biochem. J. 312, 863–870]. 4F2hc mRNA was detected in CHO cells, COS cells, activated lymphocytes isolated from mouse spleen and primary cultures of astrocytes. In all these cell types, Na+-independent isoleucine transport was mediated by system L. No contribution of system y+L to isoleucine or arginine transport was detected in C6-BU-1 cells. In lymphocytes, both system-L-like amino acid-transport activity and 4F2hc mRNA levels increased after treatment with phorbol ester plus ionomycin. Antisense oligonucleotides caused modest inhibition of Na+-independent isoleucine transport in C6-BU-1 cells and primary cultures of astroglial cells, whereas arginine transport was unaffected. Overexpression of 4F2hc cDNA in CHO cells resulted in an increase in Na+-independent isoleucine transport.

scholarly journals The 4F2hc surface antigen is necessary for expression of system L-like neutral amino acid-transport activity in C6-BU-1 rat glioma cells: evidence from expression studies in Xenopus laevis oocytes

1995 ◽  
Vol 312 (3) ◽  
pp. 863-870 ◽  
Author(s):  
S Bröer ◽  
A Bröer ◽  
B Hamprecht
Keyword(s):  
Amino Acid ◽  
Xenopus Laevis ◽  
Amino Acid Transport ◽  
Glioma Cells ◽  
Xenopus Laevis Oocytes ◽  
Acid Transport ◽  
Transport Activity ◽  
Rat Glioma ◽  
Polyadenylated Rna ◽  
System L

Mammalian cells possess a variety of amino acid-transport systems with overlapping substrate specificity. System L is one of the major amino acid-transport systems in all non-epithelial cells. Its molecular structure is not known. To clone the neutral amino acid-transporter system L, we followed an expression cloning strategy using Xenopus laevis oocytes. A cDNA library derived from C6-BU-1 rat glioma cells was used as a source, because high expression of system L activity could be demonstrated with polyadenylated RNA isolated from these cells, when injected into Xenopus laevis oocytes [Bröer, Bröer and Hamprecht (1994) Biochim. Biophys. Acta 1192, 95-100]. A single clone (ILAT) was identified, the sense cRNA of which, on injection into Xenopus laevis oocytes, stimulated sodium-independent isoleucine transport by about 100-fold. Further characterization revealed that transport of cationic amino acids was also stimulated. Sequencing of the cDNA showed that the identified clone is the heavy chain of the rat 4F2 surface antigen, a marker of tumour cells and activated lymphocytes. Uptake of neutral and cationic amino acids was not stimulated by the presence of Na+ ions. Antisense cRNA transcribed from this clone or antisense oligonucleotides, when co-injected with polyadenylated RNA from C6-BU-1 rat glioma cells, completely suppressed system L-like isoleucine-transport activity. We conclude that ILAT is necessary for expression of system L-like amino acid-transport activity by polyadenylated RNA from C6-BU-1 rat glioma cells.

scholarly journals Expression of solute carrier 7A4 (SLC7A4) in the plasma membrane is not sufficient to mediate amino acid transport activity

2002 ◽  
Vol 364 (3) ◽  
pp. 767-775 ◽  
Author(s):  
Sabine WOLF ◽  
Annette JANZEN ◽  
Nicole VÉKONY ◽  
Ursula MARTINÉ ◽  
Dennis STRAND ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Amino Acid ◽  
Protein Expression ◽  
Amino Acid Transport ◽  
Fluorescent Protein ◽  
Xenopus Laevis Oocytes ◽  
Amino Acid Transporters ◽  
Acid Transport ◽  
Transport Activity ◽  
Solute Carrier

Member 4 of human solute carrier family 7 (SLC7A4) exhibits significant sequence homology with the SLC7 subfamily of human cationic amino acid transporters (hCATs) [Sperandeo, Borsani, Incerti, Zollo, Rossi, Zuffardi, Castaldo, Taglialatela, Andria and Sebastio (1998) Genomics 49, 230–236]. It is therefore often referred to as hCAT-4 even though no convincing transport activity has been shown for this protein. We expressed SLC7A4 in Xenopus laevis oocytes, but could not detect any transport activity for cationic, neutral or anionic amino acids or for the polyamine putrescine. In addition, human glioblastoma cells stably overexpressing a fusion protein between SLC7A4 and the enhanced green fluorescent protein (EGFP) did not exhibit an increased transport activity for l-arginine. The lack of transport activity was not due to a lack of SLC7A4 protein expression in the plasma membrane, as in both cell types SLC7A4-EGFP exhibited a similar subcellular localization and level of protein expression as functional hCAT-EGFP proteins. The expression of SLC7A4 can be induced in NT2 teratocarcinoma cells by treatment with retinoic acid. However, also for this endogenously expressed SLC7A4, we could not detect any transport activity for l-arginine. Our data demonstrate that the expression of SLC7A4 in the plasma membrane is not sufficient to induce an amino acid transport activity in X. laevis oocytes or human cells. Therefore, SLC7A4 is either not an amino acid transporter or it needs additional (protein) factor(s) to be functional.

Expression and regulation of 4F2hc and hLAT1 in human trophoblasts

2002 ◽  
Vol 282 (1) ◽  
pp. C196-C204 ◽  
Author(s):  
Yoko Okamoto ◽  
Masahiro Sakata ◽  
Kazuhiro Ogura ◽  
Toshiya Yamamoto ◽  
Masaaki Yamaguchi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Line ◽  
Transport System ◽  
Human Placenta ◽  
Amino Acid Transport ◽  
Calcium Ionophore ◽  
Acid Transport ◽  
Amino Acid Transport System ◽  
System L ◽  
Choriocarcinoma Cell Line

The neutral amino acid transport system L is a sodium-independent transport system in human placenta and choriocarcinoma cells. Recently, it was found that the heterodimer composed of hLAT1 (a light-chain protein) and 4F2 heavy chain (4F2hc), a type II transmembrane glycoprotein, is responsible for system L amino acid transport. We found that the mRNAs of 4F2hc and hLAT1 were expressed in the human placenta and a human choriocarcinoma cell line. The levels of the 4F2hc and hLAT1 proteins in the human placenta increased at full term compared with those at midtrimester. Immunohistochemical data showed that these proteins were localized mainly in the placental apical membrane. Data from leucine uptake experiments, Northern blot analysis, and immunoblot analysis showed that this transport system was partially regulated by protein kinase C and calcium ionophore in the human choriocarcinoma cell line. Our results suggest that the heterodimer of 4F2hc and hLAT1 may play an important role in placental amino acid transport system L.

scholarly journals Electrogenic Na+-dependentl-alanine transport in the lizard duodenum. Involvement of systems A and ASC

2001 ◽  
Vol 280 (3) ◽  
pp. R612-R622
Author(s):  
Virtudes Medina ◽  
Antonio Lorenzo ◽  
Mario Dı́az
Keyword(s):  
Amino Acid ◽  
Amino Acid Transport ◽  
Short Circuit ◽  
Duodenal Mucosa ◽  
Neutral Amino Acid ◽  
Acid Transport ◽  
Transport Activity ◽  
System A ◽  
Alanine Transport ◽  
Gallotia Galloti

l-Alanine transport across the isolated duodenal mucosa of the lizard Gallotia galloti has been studied in Ussing chambers under short-circuit conditions. Net l-alanine fluxes, transepithelial potential difference (PD), and short-circuit current ( Isc) showed concentration-dependent relationships. Na+-dependent l-alanine transport was substantially inhibited by the analog α-methyl aminoisobutyric acid (MeAIB). Likewise, MeAIB fluxes were completely inhibited byl-alanine, indicating the presence of system A for neutral amino acid transport. System A transport activity was electrogenic and exhibited hyperbolic relationships for net MeAIB fluxes, PD, and Isc, which displayed similar apparent K m values. Na+-dependentl-alanine transport, but not MeAIB transport, was partially inhibited by l-serine and l-cysteine, indicating the participation of system ASC. This transport activity represents the major pathway for l-alanine absorption and seemed to operate in an electroneutral mode with a negligible contribution to the l-alanine-induced electrogenicity. It is concluded from the present study that the active Na+-dependent l-alanine transport across the isolated duodenal mucosa of Gallotia galloti results from the independent activity of systems A and ASC for neutral amino acid transport.

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