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Arginine transport in human erythroid cells: discrimination of CAT1 and 4F2hc/y+LAT2 roles

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Abstract

Since arginine metabolites, such as nitric oxide and polyamines, influence the expression of genes involved in erythroid differentiation, the transport of the cationic amino acid may play an important role in erythroid cells. However, available data only concern the presence in these cells of CAT1 transporter (system y+), while no information exists on the role of the heterodimeric transporters of system y+L (4F2hc/y+LAT1 and 4F2hc/y+LAT2) which operates transmembrane arginine fluxes cis-inhibited by neutral amino acids in the presence of sodium. Using erythroleukemia K562 cells and normal erythroid precursors, we demonstrate here that arginine transport in human erythroid cells is due to the additive contributions of a leucine-sensitive and leucine-insensitive component. In both cell types, leucine inhibition of arginine influx is much less evident in the absence of sodium, a hallmark of system y+L. In K562 cells, N-ethylmaleimide, a known inhibitor of CAT transporters (system y+), suppresses only a fraction of arginine influx corresponding to leucine-insensitive uptake. Moreover, in Xenopus oocytes coexpressing 4F2hc and y+LAT2, leucine exerts a marked inhibition of arginine transport, partially dependent on sodium, while no inhibition is seen in oocytes expressing CAT1. Lastly, silencing of SLC7A6, the gene for y+LAT2, lowers arginine transport and doubles the intracellular content of the cationic amino acid in K562 cells. We conclude that arginine transport in human erythroid cells is due to both system y+ (CAT1 transporter) and system y+L (4F2hc/y+LAT2 isoform), which mainly contribute, respectively, to the influx and to the efflux of the cationic amino acid.

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Abbreviations

CAA:

Cationic amino acids

CAT:

Cationic amino acid transporter

NEM:

N-Ethylmaleimide

PBS:

Phosphate-buffered saline

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Acknowledgments

Work in the laboratory of E.I.C. was supported by grant Cl 100/4-3 from the Deutsche Forschungsgemeinschaft. Work in Parma was funded by FIL, University of Parma (O.B., V.D.A., G.C.G., B.M.R.), and by MIUR PRIN 2007BY3PLL_002 “Endothelial function and inflammatory stress: role of mTORkinase” to V.D.A. R.G is granted by Fondazione CARIPARO and by Telethon (GGP07257).

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Authors and Affiliations

  1. Unit of General and Clinical Pathology, Department of Experimental Medicine, University of Parma, Via Volturno, 39-43125, Parma, Italy

    Bianca Maria Rotoli, Amelia Barilli, Rossana Visigalli, Gian C. Gazzola, Ovidio Bussolati & Valeria Dall’Asta

  2. Department of Pharmacology, Johannes-Gutenberg University, Mainz, Germany

    Ellen I. Closs, Alexandra Simon & Alice Habermeier

  3. Section of Molecular Biology, Department of Biochemistry and Molecular Biology, University of Ferrara, Ferrara, Italy

    Nicoletta Bianchi & Roberto Gambari

Authors
  1. Bianca Maria Rotoli
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  2. Ellen I. Closs
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  3. Amelia Barilli
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  4. Rossana Visigalli
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  5. Alexandra Simon
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  6. Alice Habermeier
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  7. Nicoletta Bianchi
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  8. Roberto Gambari
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  9. Gian C. Gazzola
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  10. Ovidio Bussolati
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  11. Valeria Dall’Asta
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Correspondence to Ovidio Bussolati.

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Rotoli, B.M., Closs, E.I., Barilli, A. et al. Arginine transport in human erythroid cells: discrimination of CAT1 and 4F2hc/y+LAT2 roles. Pflugers Arch - Eur J Physiol 458, 1163–1173 (2009). https://doi.org/10.1007/s00424-009-0692-9

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  • DOI: https://doi.org/10.1007/s00424-009-0692-9

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